| 1996 |
Calcium signaling induces a physical association between NF-AT4 (NFATC3) and calcineurin, and these molecules are transported as a complex to the nucleus, where calcineurin continues to dephosphorylate NF-AT4, counteracting a nuclear NF-AT kinase. |
Co-immunoprecipitation, nuclear import assays, pharmacological inhibition with cyclosporin A/FK506 |
Nature |
High |
8684469
|
| 1997 |
JNK phosphorylates NFAT4 (NFATC3) on two sites; mutational removal of these JNK phosphorylation sites causes constitutive nuclear localization of NFAT4, while JNK activation in calcineurin-stimulated cells causes nuclear exclusion of NFAT4, showing JNK opposes calcineurin-driven nuclear accumulation. |
In vitro kinase assay, site-directed mutagenesis, nuclear localization assays |
Science |
High |
9374467
|
| 1998 |
Casein kinase I alpha (CKIα) directly binds and phosphorylates NF-AT4 (NFATC3), establishing intramolecular masking of the nuclear localization signal to inhibit nuclear translocation; MEKK1 indirectly suppresses nuclear import by stabilizing the NF-AT4/CKIα interaction. |
Co-immunoprecipitation, in vitro kinase assay, nuclear translocation assays, epistasis by co-expression |
Cell |
High |
9630228
|
| 1995 |
NFATc3 (NFATx) binds NFAT sites in the IL-2 promoter when combined with AP-1 (c-Fos/c-Jun) and activates IL-2 promoter transcription; it undergoes a calcium-dependent decrease in apparent molecular mass that is inhibited by FK506, consistent with calcineurin-dependent dephosphorylation. |
EMSA, reporter gene assay in COS-7 cells, Western blot mobility shift upon calcium elevation |
Molecular and cellular biology |
Medium |
7739550
|
| 1995 |
NFATc3 activates NFAT-site-dependent transcription and exhibits a distinct DNA-binding site specificity compared to other NFATc family members, and undergoes calcium-dependent modifications blocked by FK506. |
Reporter gene assay, DNA-binding specificity assays, Western blot |
The Journal of biological chemistry |
Medium |
7650004
|
| 2000 |
Calcineurin enzymatic activity is transiently increased during initiation of myogenic differentiation in C2C12 cells and is associated with NFATc3 nuclear translocation; adenoviral calcineurin enhances differentiation while calcineurin inhibitory peptides attenuate it; NFATc3 cotransfection enhances MyoD-directed myogenesis. |
Calcineurin enzymatic assay, adenoviral gene transfer, nuclear translocation assay, MyoD co-transfection |
Molecular and cellular biology |
High |
10938134
|
| 2002 |
Targeted disruption of NFATc3 (but not NFATc4) significantly reduces calcineurin transgene-induced cardiac hypertrophy and attenuates pressure overload- and angiotensin II-induced cardiac hypertrophy, providing genetic evidence that calcineurin signaling requires NFATc3 as a downstream effector in vivo. |
Genetic knockout mice, cardiac hypertrophy models (calcineurin transgene, pressure overload, angiotensin II infusion), morphometric analysis |
Molecular and cellular biology |
High |
12370307
|
| 2004 |
NFATc3 is an obligatory component of the calcineurin-dependent signaling cascade mediating angiotensin II-induced reduction of Kv2.1 K+ channel subunit expression and decreased Kv currents in arterial smooth muscle. |
NFATc3 knockout mice, electrophysiology (Kv current recordings), Western blot, pharmacological inhibition |
The Journal of biological chemistry |
High |
15322114
|
| 2004 |
NFATc3 is required for post-myocardial infarction reductions in Kv currents (Ito, IKslow1, IKslow2) and Kv1.5, Kv2.1, Kv4.2, Kv4.3 expression; beta-adrenergic signaling activates calcineurin/NFATc3 to decrease these channels, and Kv currents do not change after MI in NFATc3 knockout mice. |
NFATc3 knockout mice, electrophysiology, Western blot, RT-PCR, calcineurin inhibitor (cyclosporine), beta-blocker treatment |
Circulation research |
High |
15087419
|
| 2006 |
NFATc3 activation down-regulates the beta1 subunit of BK channels in arterial smooth muscle during angiotensin II-induced hypertension; beta1 expression was decreased in WT but not NFATc3-null arteries, and NFATc3-null mice showed attenuated angiotensin II-induced hypertension. |
NFATc3 knockout mice, patch-clamp electrophysiology, Western blot, angiotensin II infusion model |
The Journal of biological chemistry |
High |
17148444
|
| 2006 |
Differential calcineurin/NFATc3 activity (higher in endocardial than epicardial myocytes) contributes to the transmural Ito gradient in the mouse left ventricle; NFATc3-null mice show complete loss of Ito and Kv4 heterogeneity. |
NFATc3 knockout mice, electrophysiology, NFAT reporter assay, Ca2+ imaging |
Circulation research |
High |
16614306
|
| 2007 |
NFATc3 mediates chronic hypoxia-induced pulmonary arterial remodeling; NFATc3 knockout mice do not show increased alpha-actin levels or increased arterial wall thickness after chronic hypoxia, establishing NFATc3 as required for hypoxia-induced vascular remodeling. |
NFATc3 knockout mice, NFAT-luciferase reporter mice, immunofluorescence, cyclosporin A pharmacology |
The Journal of biological chemistry |
High |
17403661
|
| 2007 |
GSK-3β suppresses myogenic differentiation through negative regulation of NFATc3 nuclear translocation and transcriptional activity; NFATc3-deficient myoblasts fail to show increased muscle gene expression following GSK-3β inhibition, establishing NFATc3 as the required downstream effector. |
GSK-3β knockout/kinase-dead mutant reconstitution, NFATc3 knockout myoblasts, nuclear translocation assay, NFAT promoter reporter, muscle gene expression |
The Journal of biological chemistry |
High |
17977834
|
| 2009 |
NFATc3 directly activates miR-23a expression through transcriptional machinery in cardiomyocytes undergoing hypertrophy; miR-23a in turn suppresses translation of MuRF1 (muscle specific ring finger protein 1), an anti-hypertrophic protein. |
Reporter gene assay (miR-23a promoter), ChIP, knockdown experiments, Western blot |
Proceedings of the National Academy of Sciences of the United States of America |
Medium |
19574461
|
| 2010 |
NFATc3 binds to the promoter region of myocardin and transcriptionally activates its expression; knockdown of myocardin attenuates hypertrophic responses triggered by NFATc3, placing myocardin downstream of NFATc3 in the hypertrophic cascade. |
ChIP, promoter reporter assay, siRNA knockdown, Western blot |
The Journal of biological chemistry |
Medium |
20177053
|
| 2001 |
Calcineurin/NFATc3 signaling is required for normal vasculogenesis; mice with disruption of both NFATc3 and NFATc4 die around E11 with generalized vascular assembly defects, and a Ca2+-insensitive calcineurin B mutation phenocopies these defects. |
Genetic double knockout mice, calcineurin B point mutation knockin, embryological analysis |
Cell |
High |
11439183
|
| 2001 |
PDGF-induced NFAT4 (NFATC3) translocation in native smooth muscle requires Ca2+ entry through voltage-dependent Ca2+ channels; elevation of [Ca2+]i by membrane depolarization or ionomycin alone is insufficient for NFAT4 nuclear accumulation, indicating Ca2+ influx is necessary but not sufficient and PDGF-induced modulation of nuclear import/export is also required. |
GFP-NFAT4 nuclear translocation assay, Ca2+ channel blockers, K+ channel opener, calcineurin blockers (CsA, FK506), luciferase reporter |
The Journal of biological chemistry |
Medium |
11278965
|
| 2002 |
NFAT4 (NFATC3) induces chondrogenesis and activates BMP2 gene expression; calcineurin/NFAT4 acts upstream of BMP signaling, since BMP antagonists block calcium/calcineurin-induced chondrogenesis. |
Calcium ionophore treatment, calcineurin inhibitor (cyclosporin A), dominant-negative BMP receptors, reporter gene assay, chondrogenic differentiation assays |
The Journal of biological chemistry |
Medium |
12239209
|
| 2003 |
Physiological intravascular pressure (100 mmHg) induces NFATc3 nuclear localization in mouse cerebral artery smooth muscle via a pathway requiring endothelial NO, cGMP-dependent kinase (PKG), voltage-dependent Ca2+ channels, and PKG-dependent inhibition of JNK2; NFATc3 nuclear accumulation is absent in JNK2-null arteries when PKG is blocked. |
NFATc3 immunofluorescence in intact arteries, endothelial denudation, pharmacological inhibitors (NOS, PKG, VDCC, JNK), JNK2 knockout mice |
The Journal of biological chemistry |
High |
14688253
|
| 2002 |
CK1 (protein kinase CK1) phosphorylates the A domain of NF-AT4 (NFATC3) via a two-phase hierarchical mechanism: an acidic linker region provides a docking site for CK1 and allows initial (unorthodox) phosphorylation of Ser177; once phosphorylated, Ser177 primes hierarchical phosphorylation of downstream serines (S180, S181, S184, S186). |
In vitro kinase assay with 27 peptide variants, site-directed mutagenesis (alanine and phosphoserine substitutions), Km/Vmax analysis |
Biochemistry |
High |
11781102
|
| 2001 |
A calcineurin-binding motif in NFATx (NFATC3) identified; a competing peptide (Pep3) based on this motif selectively blocks nuclear translocation of NFATx but not NFATp, and suppresses cytokine production by T cells expressing NFATx. |
Peptide competition assay, retrovirus-mediated gene transfer, nuclear translocation assay, cytokine measurement |
Journal of immunology |
Medium |
11509611
|
| 2008 |
PKCα-dependent persistent Ca2+ sparklets in arterial myocytes activate the calcineurin/NFATc3 signaling cascade, leading to increased Ca2+ entry, elevated arterial [Ca2+]i, and enhanced myogenic tone during hypertension; PKCα ablation is protective against angiotensin II-induced hypertension. |
Total internal reflection fluorescence microscopy (Ca2+ sparklet imaging), PKCα knockout mice, NFATc3 reporter assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
18832165
|
| 2008 |
NFATc3 regulates BK channel alpha-subunit expression in urinary bladder smooth muscle; NFATc3-null mice show reduced BK currents, decreased BK alpha-subunit mRNA, and elevated contractile responses to electrical field stimulation. |
NFATc3 knockout mice, patch-clamp electrophysiology, RT-PCR, contractility assay |
American journal of physiology. Cell physiology |
High |
18579799
|
| 2009 |
NFATc3 is required for chronic hypoxia-induced upregulation of soluble guanylyl cyclase alpha1 (sGC-α1) expression in pulmonary arterial smooth muscle; NFATc3 binds functional sites in the sGC-α1 promoter and activates its transcription. |
NFATc3 knockout mice, cyclosporin-treated mice, promoter reporter assay, NFAT binding site identification |
American journal of physiology. Lung cellular and molecular physiology |
Medium |
19592461
|
| 2011 |
Endothelin-1 (ET-1) contributes to chronic hypoxia-induced NFATc3 activation in pulmonary arteries via RhoA/Rho kinase (ROK) and calcineurin; ROK increases actin polymerization to provide structural support for NFATc3 nuclear transport, downstream of Ca2+-calcineurin-dependent dephosphorylation. |
NFAT-luciferase reporter mice, pharmacological inhibitors (ET receptor, calcineurin, L-type Ca2+ channels, ROK), GFP-NFATc3 nuclear import assay in human PASMC |
American journal of physiology. Cell physiology |
Medium |
21525433
|
| 2011 |
NFATc3 is specifically required for IL-2 and COX-2 gene expression in T cells and for T-cell proliferation; NFATc3 also regulates COX-2 in endothelial cells where it is required for COX-2-dependent migration and angiogenesis in vivo. |
siRNA knockdown, reporter gene assay, cell migration assay, in vivo angiogenesis assay |
Blood |
Medium |
21642596
|
| 2011 |
NFATc3 directly binds to the RANKL promoter and stimulates RANKL expression in osteoblasts in response to high extracellular calcium; NFATc1 acts upstream by inducing NFATc3 expression and activity. |
ChIP, reporter assay, siRNA knockdown, overexpression |
Bone |
Medium |
21514407
|
| 2012 |
In neurons, NFATc3 undergoes rapid dephosphorylation and nuclear translocation (complete within 20 min) upon Ca2+ elevation, while NFATc4 requires prolonged depolarization; the serine-proline repeat region of NFATc3 is critical for the magnitude of nuclear accumulation; knockdown of NFATc3 strongly diminishes NFAT-mediated transcription induced by mild depolarization. |
GFP-NFAT chimera nuclear translocation assay, NFATc3/c4 siRNA knockdown, phosphorylation immunoblot, luciferase reporter, domain-swap chimeras |
The Journal of biological chemistry |
Medium |
22977251
|
| 2012 |
NFATc3 is expressed in pancreatic acinar cells and regulates trypsinogen activation and inflammatory cytokine (CXCL2) expression; NFATc3-deficient mice are protected from taurocholate- and L-arginine-induced acute pancreatitis. |
NFATc3 knockout mice, NFAT-luciferase reporter mice, NFAT inhibitor (A-285222), confocal immunofluorescence, trypsin activity assay |
Gastroenterology |
High |
22841788
|
| 2013 |
NFATc3 binds to the RANTES promoter and directly regulates RANTES gene transcription in rheumatoid arthritis synovial fibroblasts in response to ASIC1a-mediated Ca2+ influx. |
ChIP-qPCR, dual-luciferase reporter assay, calcium imaging, ASIC1a overexpression/knockdown |
Theranostics |
Medium |
31903118
|
| 2014 |
Trim17 binds preferentially SUMOylated forms of NFATc3 and reduces its calcium-mediated nuclear localization by approximately 2-fold without promoting its ubiquitination/degradation; NFATc3 induces transcription of the proapoptotic gene Trim17 via binding to its promoter together with c-Jun, creating a feedback loop. |
Co-immunoprecipitation, luciferase reporter, nuclear translocation assay, ChIP, siRNA knockdown, neuronal apoptosis assay |
Cell death and differentiation |
Medium |
25215946
|
| 2014 |
T. gondii dense granule protein GRA6 selectively activates NFAT4 (NFATC3) via CAMLG (calcium modulating ligand); GRA6-deficient parasites fail to activate NFAT4, and NFAT4-deficient mice show decreased chemokine (Cxcl2, Ccl2) expression and altered immune cell recruitment at infection sites. |
GFP-NFAT4 reporter assay, GRA6 knockout parasites, NFAT4 knockout mice, overexpression, NFAT inhibitor treatment |
The Journal of experimental medicine |
High |
25225460
|
| 2015 |
NFATc3 inhibits miR-324-5p expression; miR-324-5p suppresses translation of Mtfr1 (mitochondrial fission regulator 1); knockdown of NFATc3 suppresses mitochondrial fission, cardiomyocyte apoptosis and myocardial infarction. |
ChIP, luciferase reporter, siRNA knockdown, mitochondrial morphology imaging, apoptosis assays |
Cell death & disease |
Medium |
26633713
|
| 2016 |
NFATC3 specifically binds to IRF7 protein and enhances IRF7-mediated type I IFN production; NFATC3 knockout greatly reduces CpG DNA-induced nuclear translocation of IRF7; both NFATC3 and IRF7 bind to type I IFN promoters and the NFAT binding site in IFN promoters is required for IRF7-mediated IFN expression. |
Co-immunoprecipitation, NFATC3 knockout (in vitro and in vivo), ChIP, reporter gene assay, NFAT binding site mutagenesis |
The Journal of experimental medicine |
High |
27697837
|
| 2016 |
ASIC1-mediated Ca2+ influx promotes NFATc3 nuclear import in pulmonary arterial smooth muscle cells via a PICK1 scaffold protein that interacts with both ASIC1 and calcineurin; PICK1 inhibition abolishes NFATc3 nuclear import without altering Ca2+ responses. |
ASIC1 knockout mice, pharmacological ASIC1 inhibition, Duolink proximity ligation assay, PICK1 inhibitor, GFP-NFATc3 nuclear import assay |
American journal of physiology. Lung cellular and molecular physiology |
High |
27190058
|
| 2017 |
NFATc3 directly targets the Etv2 promoter via an evolutionarily conserved cis-element to drive hematopoietic fate commitment in embryonic stem cells; constitutively active NFATc3 rescues Etv2 expression and hematopoietic progenitor formation in IP3R-triple-knockout cells. |
IP3R triple-knockout embryonic stem cells, constitutively active NFATc3 overexpression rescue, ChIP showing NFATc3 binding to Etv2 promoter, NFAT reporter assay |
Journal of molecular cell biology |
Medium |
28419336
|
| 2019 |
NFATc3 is SUMOylated; hypoxia-induced SENP3-mediated deSUMOylation of NFATc3 at K384 impairs the interaction between NFATc3 and GSK-3β, decreases NFATc3 phosphorylation, and increases NFATc3 nuclear occupancy to promote pancreatic carcinoma progression. |
SUMOylation site mutation (K384), co-immunoprecipitation, SENP3 knockdown, nuclear fractionation, in vitro deSUMOylation |
Cell death & disease |
Medium |
35484132
|
| 2019 |
CHIP (carboxyl-terminus of Hsc70-interacting protein, an E3 ubiquitin ligase) promotes proteasomal degradation of NFATc3; co-immunoprecipitation demonstrates physical interaction between CHIP and NFATc3, and CHIP overexpression reduces NFATc3 protein levels and attenuates hypertrophy and apoptosis. |
Co-immunoprecipitation, Western blot, proteasome inhibitor treatment, CHIP overexpression |
Journal of cellular physiology |
Medium |
30980393
|
| 2020 |
NFATc3 transcriptionally upregulates miR-204 in macrophages; miR-204-5p suppresses SR-A expression via canonical 3'UTR targeting, while nuclear miR-204-3p inhibits CD36 transcription; this NFATc3/miR-204 axis prevents foam cell formation and atherosclerosis. |
Macrophage-specific NFATc3 knockout and transgenic mice, ChIP, miR-204 rescue experiments, luciferase reporter |
European heart journal |
High |
34570211
|
| 2020 |
NFATc3 activates miR-153-3p expression in cardiomyocytes; ChIP-qPCR and luciferase reporter assay confirmed NFATc3 as upstream transcriptional regulator of miR-153-3p; miR-153-3p suppresses Mfn1 translation, promoting mitochondrial fission and hypertrophy. |
ChIP-qPCR, luciferase reporter assay, siRNA knockdown, mitochondrial morphology imaging |
Theranostics |
Medium |
31903137
|
| 2021 |
NFATc3 binds to the promoters of IFNL1 and IFNB1 genes in hepatocytes to promote IFN production; RIG-I pathway activation increases NFATc3 nuclear localization, which further enhances RIG-I-mediated IFN responses (positive feedback). |
ChIP, reporter gene assay, gain- and loss-of-function experiments, nuclear localization assay |
Oncoimmunology |
Medium |
33520407
|
| 2022 |
Trim39 is an E3 ubiquitin ligase for NFATc3 that acts preferentially on SUMOylated forms of NFATc3; Trim39 binds and ubiquitinates NFATc3 in vitro and in cells, reducing NFATc3 protein level and transcriptional activity; Trim17 inhibits Trim39-mediated ubiquitination of NFATc3 by reducing both Trim39 E3 activity and the NFATc3/Trim39 interaction. |
In vitro ubiquitination assay, co-immunoprecipitation, siRNA knockdown, luciferase reporter, SUMOylation site mutagenesis, SUMO-interacting motif mutagenesis |
Cell death and differentiation |
High |
35449213
|
| 2013 |
NFATc3 promotes Ca2+-dependent MMP3 expression in astrocytes; constitutively active NFATc3 induces Mmp3 expression, and a Mmp3 promoter luciferase reporter is activated by increased [Ca2+]i via calcineurin/NFAT, while other MMPs are unaffected. |
Constitutively active NFATc3 overexpression, calcium ionophore, cyclosporin A inhibition, promoter reporter assay, in vivo stab-wound model |
Glia |
Medium |
23625833
|
| 2013 |
NFATc3 binds to the -GGAAA- sequence in the COX-2 promoter and transcriptionally activates COX-2 expression in cardiomyocytes in response to endothelin-1, as demonstrated by ChIP and promoter reporter assay. |
ChIP, promoter reporter assay, NFATc3 knockdown, cyclosporin A |
Molecular and cellular endocrinology |
Medium |
24291639
|
| 2014 |
Nuclear Raf-1 phosphorylated at Ser621 associates with NFATc3 near its cognate binding site in the CXCR5 promoter, driving transcriptional upregulation of CXCR5 during retinoic acid-induced differentiation of HL-60 leukemic cells. |
Co-immunoprecipitation, ChIP, pharmacological inhibition (PD98059), reporter assay |
The FEBS journal |
Medium |
24330068
|
| 2021 |
NFATc3 regulates CXCL2 gene expression in macrophages (CXCL2 promoter-luciferase reporter activity is accentuated by co-transfection with NFATc3 expression vector); NFATc3-deficient macrophages fail to produce CCL2 and CXCL2 in response to IL-33 or conditioned medium from bleomycin-treated epithelial cells, and adoptive transfer of NFATc3+/+ macrophages restores susceptibility to pulmonary fibrosis in NFATc3+/- mice. |
Promoter-luciferase reporter assay, macrophage-specific conditional KO, adoptive transfer, in vitro cytokine stimulation |
Aging and disease |
Medium |
37523510
|
| 2017 |
NFATc3 occupies the RAG1 promoter for transcriptional repression; NFATc3 knockdown increases RAG1 expression and double-strand DNA damage in gastric cancer cells, while NFATc3 overexpression blocks RAG1 expression and DNA damage induced by arsenic sulfide. |
ChIP-qPCR, RNA-seq, siRNA knockdown, lentiviral overexpression, Western blot |
Journal of experimental & clinical cancer research |
Medium |
31822296
|
| 2019 |
NFATc3 binds to the OCT4 promoter and activates OCT4 transcription to promote cancer stem cell self-renewal in oral squamous cell carcinoma; inhibition of OCT4 abrogates the CSC phenotype driven by NFATc3. |
ChIP, promoter reporter assay, NFATc3 overexpression and knockdown, OCT4 rescue experiments, tumor sphere formation assay |
Oncotarget |
Medium |
31040921
|
| 2017 |
NFATc3 promotes myocardin (and AQP2) promoter activity; NFATc3 knockout mice display symptoms of nephrogenic diabetes insipidus (polyuria, reduced AQP2 expression) and ILK-conditional-knockdown mice show reduced nuclear localization of NFATc3, linking ILK→GSK3β→NFATc3 to AQP2 transcription. |
NFATc3 knockout mice, ILK conditional knockdown, reporter gene assay, nuclear localization assay |
Biochimica et biophysica acta. Gene regulatory mechanisms |
Medium |
28736155
|
| 2021 |
NFATc3 interacts with c-JUN and regulates the expression of SRPX2; SRPX2 knockdown diminishes the upregulation of lineage and EMT markers induced by co-overexpression of NFATc3 and c-JUN in human embryonic stem cells, placing SRPX2 downstream of the NFATc3/c-JUN complex. |
Co-immunoprecipitation (NFATc3/c-JUN), siRNA knockdown, overexpression, differentiation marker assays |
Journal of cellular physiology |
Medium |
33393109
|
| 2006 |
Activity-independent nucleocytoplasmic shuttling of NFATc1 occurs in resting skeletal muscle fibers (leptomycin B causes nuclear accumulation); however, NFATc3 does NOT show similar shuttling in resting fibers — leptomycin B did not cause nuclear accumulation of NFATc3 in unstimulated fibers, demonstrating isoform-specific regulation. |
Live-cell imaging of GFP-NFAT isoforms in adult skeletal muscle fibers, leptomycin B treatment, electrical stimulation, calcineurin inhibition |
Molecular biology of the cell |
Medium |
16436503
|