| 2001 |
Targeted deletion of exon 5 (eliminating the first Ig domain) of Dutt1/Robo1 in mice causes neonatal respiratory failure due to delayed lung maturation (reduced air spaces, increased mesenchyme) and bronchial epithelial hyperplasia in survivors, establishing a direct functional role for Robo1 in lung development. |
Targeted gene knockout in mice (homozygous deletion of exon 5); histopathology and phenotypic analysis |
Proceedings of the National Academy of Sciences of the United States of America |
High |
11734623
|
| 2004 |
Dutt1/Robo1 heterozygous mice spontaneously develop lymphomas and lung adenocarcinomas; tumors retain the structurally normal allele but show promoter methylation and reduced Robo1 protein, consistent with Knudson two-hit tumor suppressor mechanism. |
Heterozygous knockout mouse model; LOH analysis; bisulfite sequencing of promoter; Western blot and immunohistochemistry for protein expression |
Cancer research |
High |
15374951
|
| 2002 |
The DUTT1/ROBO1 promoter CpG island is hypermethylated in breast, lung, and renal cell carcinoma tumors; methylation silences expression, which is reactivated by 5-aza-2'-deoxycytidine treatment, identifying promoter methylation as an epigenetic inactivation mechanism. |
Bisulfite sequencing; methylation-specific PCR; RT-PCR expression analysis; 5-aza-2'-deoxycytidine demethylation assay |
Oncogene |
High |
12082532
|
| 2006 |
Robo1 knockout mice display major forebrain axon pathfinding defects (corpus callosum and hippocampal commissure dysgenesis, premature corticothalamic/thalamocortical targeting) and up to twice as many cortical interneurons migrate into the cortex, establishing Robo1 as a regulator of both major axon tract formation and interneuron migration. |
Robo1 knockout mice (targeted deletion of exon 5); MRI diffusion tensor imaging; immunohistochemistry; cell counting |
Development (Cambridge, England) |
High |
16690755
|
| 2007 |
RNAi-mediated knockdown of Robo1 in glioma cell lines abolishes the repulsive/migratory-inhibitory effect of recombinant Slit2, demonstrating that Robo1 is the major functional receptor for Slit2-mediated chemorepulsion in glioma cells. |
siRNA knockdown of Robo1; modified Boyden chamber migration assay with recombinant Slit2 |
Journal of neuro-oncology |
Medium |
17968499
|
| 2007 |
The C. elegans kinesin-related protein VAB-8L promotes posterior cell/growth-cone migration by increasing surface levels of the Robo1 ortholog SAX-3; the Rac GEF UNC-73 (Trio) physically interacts with both VAB-8L and the intracellular domain of SAX-3, supporting a model in which receptor surface localization is regulated by this complex. |
C. elegans genetics (epistasis); co-immunoprecipitation; receptor abundance measurement; gain/loss-of-function assays |
Nature neuroscience |
High |
17237778
|
| 2008 |
Robo1 forms a heterodimeric complex with Robo4; Robo1 is required for Robo4-mediated filopodia induction in endothelial cells; the Robo4 intracellular domain binds WASP, N-WASP, and WIP (actin nucleation-promoting factors) as identified by yeast two-hybrid and GST pulldown, linking Robo1/Robo4 signaling to actin-based cell motility. |
siRNA knockdown; Robo4-GFP overexpression; yeast two-hybrid; GST pulldown; endothelial tube formation and migration assays; co-immunoprecipitation |
FASEB journal |
High |
18948384
|
| 2010 |
Robo1 and Robo2 have distinct and collaborative roles in post-crossing commissural axon guidance: Robo2 is required for axons to project away from the floor plate into the lateral funiculus, while Robo1 prevents axonal stalling after midline crossing; genetic epistasis with Robo3 mutants further defines their collaborative role in preventing midline re-entry. |
Genetic combination mutants (Robo1, Robo2, Robo3 single and double/triple knockouts); axon trajectory analysis in mouse spinal cord |
The Journal of neuroscience |
High |
20631173
|
| 2010 |
Human ROBO1 undergoes sequential proteolytic cleavage by metalloproteinases (producing ROBO1-CTF1 at 129 kDa) followed by gamma-secretase cleavage (producing ROBO1-CTF2 at 118 kDa); the resulting intracellular fragment accumulates in the nucleus in cancer cells, and nuclear accumulation is abolished by either metalloproteinase inhibitor TAPI-1 or gamma-secretase inhibitor L-685,458. |
Western blot of cleavage fragments; pharmacological inhibition with TAPI-1 and L-685,458; nuclear fractionation; determination of precise cleavage site |
FEBS letters |
Medium |
20471383
|
| 2011 |
SLIT/ROBO1 signaling in the mammary basal cell layer restricts basal cell proliferation by inhibiting canonical WNT signaling, shifting β-catenin from the nuclear pool to cytoplasmic/membrane pools; loss of SLIT/ROBO1 signaling specifically in basal cells results in excess basal cells and precocious branching morphogenesis. TGF-β1 acts upstream of Robo1, inducing its expression in the basal layer. |
Conditional loss-of-function mouse models; mammary gland whole-mount analysis; immunofluorescence for β-catenin localization; Western blot; TGF-β1 treatment |
Developmental cell |
High |
21664580
|
| 2013 |
Slit2-Robo1 signaling inhibits Cdc42 activity in glioma cells, and this is the mechanism by which the pathway suppresses glioma cell invasion; Cdc42-GTP inactivation mediates the anti-invasive effect. |
Small GTPase pull-down assay (Cdc42 activity); siRNA knockdown; migration/invasion assay |
Neuroscience bulletin |
Low |
21113198
|
| 2013 |
Heparin binds to Robo1 with an affinity of ~650 nM; N-sulfo and 6-O-sulfo groups on heparin are essential for the Robo1-heparin interaction; full-length heparin chains and higher sulfation levels increase binding affinity. |
Surface plasmon resonance (SPR); chemical modification of heparin; SPR solution competition studies |
Biochimie |
Medium |
23994753
|
| 2014 |
FLRT3 is a novel co-receptor for Robo1; in thalamic axons, Slit1 acting through both Robo1 and FLRT3 induces Netrin-1 attraction by upregulating surface DCC via activation of protein kinase A (PKA); thalamic axons lacking FLRT3 are insensitive to Netrin-1, and loss of FLRT3 causes axon guidance defects in vivo. |
Co-receptor identification; in vitro axon guidance assays; PKA activation assays; surface DCC measurement; in vivo guidance phenotype analysis in knockout mice |
Current biology : CB |
High |
24560577
|
| 2014 |
ROBO1 negatively regulates cell motility and invasiveness of prostate cancer cells through interaction with DOCK1 (via its SH3-SH2 domain), controlling Rac1 activation; loss of ROBO1 leads to Rac1 activation, which destabilizes E-cadherin/β-catenin cytoskeleton and induces cell migration. |
ROBO1-expressing and ROBO1-C2C3 mutant constructs; co-immunoprecipitation; confocal microscopy; luciferase reporter; migration/invasion assays |
International journal of cancer |
Medium |
24752651
|
| 2014 |
Robo1 knockout in the developing neocortex causes a significant increase in progenitor cells and both early- and late-born pyramidal neuron populations due to prolonged proliferative activity of progenitors; confirmed by in utero electroporation of Robo1 shRNA and in dissociated cortical cell cultures. |
Robo1 knockout mice; in utero electroporation with shRNA; dissociated cortical cell cultures; BrdU/Ki67 proliferation analysis; layer-specific marker immunostaining |
The Journal of neuroscience |
High |
24741061
|
| 2015 |
Slit2 acting through Robo1 and Robo2 promotes endothelial cell migration and retinal angiogenesis; Robo1 and Robo2 are required for both Slit2- and VEGF-induced Rac1 activation and lamellipodia formation in retinal endothelial cells. |
Conditional knockout mice (endothelial-specific deletion of Slit2, Robo1, Robo2); mouse postnatal retinal angiogenesis assay; Rac1 activation assay; lamellipodia measurement; ocular neovascular disease model |
Nature medicine |
High |
25894826
|
| 2015 |
Two distinct heparin-binding sites on Robo1 were identified: one at the previously known site and a second at the disordered N-terminus; mutagenesis of the N-terminal site decreased heparin binding affinity; heparin binding also induces a minor conformational change in the Ig2 domain. |
Electron transfer dissociation-based high spatial resolution hydroxyl radical protein footprinting; surface plasmon resonance; heparin affinity chromatography; site-directed mutagenesis |
The Journal of biological chemistry |
High |
25752613
|
| 2015 |
SLIT2/ROBO1 signaling regulates mammary stem cell self-renewal by governing the choice between asymmetric and symmetric cell divisions through the transcription factor SNAI1 and spindle orientation protein mINSC; loss of SLIT2/ROBO1 increases nuclear SNAI1, which upregulates mInsc, increasing symmetric divisions and expanding the stem cell pool. |
Conditional knockout mouse model; single-cell lineage tracing; mInsc/SNAI1 expression analysis; SLIT2 treatment rescue experiments; mammary gland outgrowth assay |
Cell reports |
High |
26440891
|
| 2015 |
Activation of Slit2-Robo1 signaling promotes liver fibrosis by activating hepatic stellate cells (HSCs); mechanistically, Robo1 signaling phosphorylates Smad2, Smad3, PI3K, and AKT independently of TGF-β1, and promotes α-SMA expression; Robo1 neutralizing antibody R5 and Robo1 siRNA block these effects in vitro and attenuate CCl4-induced fibrosis in vivo. |
Slit2 transgenic and Robo1/2 heterozygous knockout mice; CCl4-induced fibrosis model; Robo1 siRNA and neutralizing antibody (R5); Western blot for Smad2/3, PI3K, AKT, α-SMA phosphorylation |
Journal of hepatology |
High |
26264936
|
| 2015 |
Slit2/Robo1 signaling promotes intestinal tumorigenesis through Src-mediated activation of the Wnt/β-catenin pathway; mechanistically, Robo1 activation by Slit2 activates Src, which downregulates E-cadherin, thereby releasing β-catenin to activate Wnt target genes. |
Slit2 transgenic and ApcMin/+ mouse models; DMH/DSS colorectal carcinoma model; Western blot; immunohistochemistry; pharmacological Src inhibition |
Oncotarget |
Medium |
25605242
|
| 2015 |
srGAP1 is a Robo1-interacting protein that mediates the anti-migratory function of Slit2-Robo1 in colorectal cancer cells by inhibiting Cdc42 activity; co-immunoprecipitation confirmed the Robo1-srGAP1 interaction; immunofluorescence showed similar dynamic subcellular distribution upon Slit2 treatment. |
Co-immunoprecipitation; immunofluorescence; Cdc42 GTPase pull-down assay; wound healing migration assay |
Journal of experimental & clinical cancer research |
Medium |
27923383
|
| 2015 |
Recombinant Slit2 reduces peripheral immune cell infiltration after surgical brain injury through Robo1-srGAP1-mediated inhibition of Cdc42 activity; decoy Robo1 co-administration and srGAP1 siRNA reversed the protective effects of Slit2, establishing the Robo1-srGAP1-Cdc42 signaling axis in immune cell migration. |
Rat surgical brain injury model; recombinant Slit2 and decoy Robo1; srGAP1 siRNA; Cdc42 activity assay; brain edema measurement; immunohistochemistry |
Neurobiology of disease |
Medium |
26550694
|
| 2015 |
Slit2 promotes angiogenesis via the Robo1-VEGFR2-ERK1/2 pathway; Slit2 overexpression upregulates Robo1, VEGFR2, and phosphorylated ERK1/2 in both transgenic mice and HUVECs, and promotes endothelial cell proliferation, migration, and tube formation. |
Slit2 transgenic mice; laser-induced CNV model; HUVEC transfection; Western blot for VEGFR2 and p-ERK1/2; proliferation, migration and tube formation assays |
Investigative ophthalmology & visual science |
Medium |
26244297
|
| 2016 |
Heparan sulfate octasaccharide ligand requirements for Robo1 were defined: Robo1 preferentially binds structures with 6-O-sulfate esters (critical), whereas 2-O-sulfate esters substantially reduce binding; a high-affinity HS octasaccharide ligand was synthesized and confirmed by SPR to bind Robo1 and reverse Slit2-Robo1-mediated inhibition of endothelial cell migration. |
HS affinity enrichment; HILIC-MS/MS structural determination; chemical synthesis of HS oligosaccharides; surface plasmon resonance (SPR) binding studies; endothelial cell migration assay |
Journal of the American Chemical Society |
High |
27611601
|
| 2016 |
NMR characterization of Robo1-Ig1-2 interaction with a synthetic heparan sulfate tetramer identified K81 as part of the binding site (Kd ~255 µM); saturation transfer difference and trNOE defined the binding epitope and bound conformation of HS; paramagnetic perturbation localized the binding site; a structural model of the complex was generated. |
Single amino acid type NMR labeling; STD NMR; trNOE; paramagnetic perturbation; complex modeling |
ACS chemical biology |
High |
27653286
|
| 2016 |
A stiff ECM increases Robo1 levels by downregulating miR-203; SLIT2/ROBO1 signaling stimulates Rac activity and focal adhesion kinase (FAK) to enhance cell tension, maintain cell shape, and facilitate matrix adhesion; Robo1 ablation in the mammary gland compromises actin stress fiber assembly and inhibits cell contractility. |
miR-203 and Robo1 manipulation; Rac activation assay; FAK activity assay; actin staining; mammary gland conditional knockout; tunable hydrogel substrates |
The Journal of cell biology |
Medium |
26975850
|
| 2018 |
Crystal structure of Robo1 Ig1-4 and negative-stain EM reconstruction of the full ectodomain show that Robo1 forms compact dimers mediated by central Ig domains, which further interact in a 'back-to-back' fashion to generate tetrameric assemblies; no change in oligomerization is observed upon Slit2-N binding, suggesting Slit2-N induces a conformational change rather than oligomerization change. |
X-ray crystallography (Robo1 Ig1-4 and Ig5); negative stain electron microscopy; fluorescent imaging with dimeric Slit2-N |
Structure (London, England : 1993) |
High |
29307485
|
| 2018 |
SLIT2/ROBO1 axis promotes the Warburg effect (aerobic glycolysis) in osteosarcoma cells via activation of the SRC/ERK/c-MYC/PFKFB2 signaling pathway; knockdown or overexpression experiments in vitro and in vivo confirmed this metabolic-oncogenic role. |
siRNA knockdown and overexpression; xenograft mouse model; Western blot for SRC, ERK, c-MYC, PFKFB2; metabolic assays |
Cell death & disease |
Medium |
29523788
|
| 2018 |
USP33 (ubiquitin-specific protease 33) deubiquitinates and stabilizes Robo1; reduced USP33 in gastric cancer leads to increased Robo1 ubiquitination and degradation, diminishing the inhibitory effect of Slit2-Robo1 signaling on cell migration and EMT. |
Co-immunoprecipitation; ubiquitination protein analysis; siRNA knockdown; in vitro and in vivo migration/EMT assays |
Cell proliferation |
Medium |
30896071
|
| 2018 |
miR-92 suppresses Robo1 translation (without mRNA degradation) in commissural neurons by binding the Robo1 3'UTR; disrupting miR-92/Robo1 interaction induces premature Slit2 repulsion and causes commissural axon projection defects in vivo; this mechanism regulates the temporal control of Slit sensitivity during midline crossing. |
miR-92 overexpression and inhibition; luciferase 3'UTR reporter assay; in vitro Slit2 repulsion assay; in vivo commissural axon labeling in chick embryo |
Cell reports |
High |
30184503
|
| 2019 |
Alternative splicing at a conserved microexon in mammalian ROBO1 and ROBO2, regulated by NOVA splicing factors, generates isoforms with distinct guidance activities; precrossing ROBO1 isoforms are inhibited to allow midline crossing, while postcrossing disinhibition prevents midline re-entry; this provides temporal regulation of Robo-mediated axonal repulsion independent of protein expression levels. |
Alternative splicing analysis; NOVA factor manipulation; in vitro Slit repulsion assay; commissural axon trajectory analysis in vivo |
eLife |
High |
31392959
|
| 2019 |
Slit2/Robo1 signaling promotes tumor metastasis in colorectal carcinoma partly through activation of the TGF-β/Smads pathway; blocking Slit2-Robo1 binding inactivates TGF-β/Smads and inhibits tumor cell migration, which can be partially restored by exogenous TGF-β1. |
Robo1 blocking antibody; TGF-β1 rescue experiment; Smad phosphorylation Western blot; migration assay; in vivo tumor models |
Cells |
Medium |
31242633
|
| 2020 |
SLIT2 inhibits macropinocytosis in macrophages and RAS-transformed cancer cells through Robo1; mechanistically, SLIT2/ROBO1 signaling induces cytoskeletal changes that oppose cortical cytoskeletal remodeling required for macropinocytosis; in vivo, SLIT2 attenuates muramyl dipeptide uptake, preventing NOD2-dependent NF-κB activation and CXCL1 secretion. |
In vitro macropinocytosis assays; in vivo mouse model with SLIT2 treatment; blocking endogenous SLIT2; cytoskeletal imaging; NOD2/NF-κB pathway analysis |
Nature communications |
High |
32807784
|
| 2020 |
PRRG4 promotes breast cancer metastasis by recruiting E3 ubiquitin ligase NEDD4 (through LPSY and PPPY motifs) to ubiquitinate and degrade Robo1, thereby activating Src and FAK signaling and promoting invasion. |
Co-immunoprecipitation; ubiquitination assay; PRRG4 knockdown/overexpression; experimental metastasis model; Western blot for Src/FAK phosphorylation |
Oncogene |
Medium |
33037408
|
| 2010 |
Src kinase activates Abl kinase, which stabilizes Robo1 protein to promote cell migration; inhibition of Abl by siRNA or kinase inhibitors decreases Robo1 protein levels and suppresses migration of transformed cells; Robo1 utilizes Cdc42 and Rac1 GTPases downstream to induce cell migration. |
Abl siRNA knockdown; pharmacological kinase inhibitors; Western blot for Robo1 protein levels; migration assay; small GTPase activation assay |
Oncotarget |
Medium |
21301049
|
| 2012 |
Slit2N/Robo1 inhibits HIV-gp120-induced migration and podosome formation in immature dendritic cells by causing Robo1 to sequester WASp and LSP1 away from β-actin, preventing WASp-Arp2/3-β-actin complex formation; Slit2N also inhibits Src signaling and downstream Rac1, Pyk2, paxillin, and CDC42 activation. |
Co-immunoprecipitation; transendothelial migration assay; podosome formation assay; Rac1/CDC42 activation assay; Src signaling Western blot |
PloS one |
Medium |
23119100
|
| 2015 |
Slit2/Robo1 signaling is required for Drosophila Robo1 midline repulsion; Ig1 domain deletion disrupts Slit binding in vitro and abolishes ectopic midline repulsion in gain-of-function studies; Ig1 is not required for proper Robo1 expression, axonal localization, or Commissureless-dependent regulation in vivo. |
In vitro Slit binding assay in Drosophila cells; Drosophila in vivo gain-of-function and genetic rescue assays; immunostaining for axonal localization and Comm-dependent regulation |
G3 (Bethesda, Md.) |
High |
26362767
|
| 2018 |
Fn3 domain deletion in Drosophila Robo1 impairs exclusion of Robo1 from commissures and reduces Commissureless-dependent downregulation of Robo1, while Fn1 and Fn2 deletions do not affect Slit binding, expression, midline repulsive function, or axonal localization; all three Fn repeats are individually dispensable for midline repulsion itself. |
In vivo domain deletion analysis (Fn1, Fn2, Fn3); in vitro Slit binding assay; midline repulsion genetic rescue assay; immunostaining for localization and Comm-dependent regulation |
G3 (Bethesda, Md.) |
Medium |
29217730
|
| 2015 |
Robo1 promotes angiogenesis in hepatocellular carcinoma by promoting Cdc42 expression in endothelial cells; knockdown of Robo1 distorts the actin cytoskeleton in HUVECs; in vivo, overexpression of Robo1 in endothelial cells mixed with hepatoma cells enhances tumor growth and microvessel density. |
Robo1 overexpression and knockdown in HUVECs; nude mouse tumor implantation; Cdc42 expression analysis; actin cytoskeleton imaging |
Tumour biology |
Medium |
26022159
|
| 2015 |
Slit2/Robo1 signaling inhibits HUVEC migration in a Robo4-dependent manner; Robo4 negatively regulates the Slit2-Robo1 migratory signal through an intracellular mechanism; CdGAP (a Rac1/Cdc42-specific GTPase activating protein) was identified by targeted proteomics as a Robo1-interacting protein and co-immunoprecipitated with Robo1. |
Boyden chamber migration assay; Robo1 and Robo4 siRNA knockdown; Robo1 extracellular domain blocking; targeted proteomics; co-immunoprecipitation of Robo1-CdGAP |
Biochemical and biophysical research communications |
Medium |
26713366
|
| 2021 |
YY1 transcription factor directly binds the ROBO1 promoter to promote transcription, as demonstrated by luciferase reporter, ChIP, and EMSA assays; ROBO1 inhibits pancreatic cancer cell proliferation through the CCNA2/CDK2 axis, causing S-phase arrest. |
Luciferase reporter gene assay; chromatin immunoprecipitation (ChIP); electrophoretic mobility shift assay (EMSA); cell cycle analysis; subcutaneous xenograft in nude mice |
Oncogene |
High |
33714986
|
| 2022 |
Glial-secreted Netrins regulate Drosophila Robo1 signaling threshold levels in neural stem cells; Netrin-Frazzled/DCC signaling modulates Robo1 levels through Abelson kinase; unbalanced Robo1 signaling impairs asymmetric stem cell division by dysregulating aPKC, Canoe/Afadin, and Numb through Rac1 and Cdc42. |
Drosophila genetics (Netrin, Frazzled/DCC, Abelson kinase, Robo1 manipulations); immunostaining for ASCD machinery components; larval brain neuroblast lineage analysis |
Current biology : CB |
Medium |
35472309
|
| 2023 |
ROBO1 exhibits dependence receptor (DR) characteristics in pancreatic ductal adenocarcinoma liver metastasis; PTP4A2 (protein tyrosine phosphatase 4A2) dephosphorylates ROBO1 in recurrent glioblastoma to modulate its downstream signaling and tumorigenicity; genetic or pharmacological inhibition of PTP4A2 acts through ROBO1; ROBO1-targeted CAR T cells eradicate tumors in xenograft models. |
CRISPR genetic perturbation screens; PTP4A2 inhibition; ROBO1 dephosphorylation assay; CDX xenograft models; CAR T cell therapy |
Nature medicine |
Medium |
39095594
|
| 2023 |
SLIT2-ROBO1-mediated coadaptation facilitates implantation and outgrowth of pancreatic ductal adenocarcinoma disseminated tumor cells in the liver premetastatic niche; ROBO1 displays dependence receptor characteristics; a ROBO1-neutralizing antibody significantly attenuates liver metastasis. |
Mouse premetastatic/macrometastatic niche models; CT/3D bioluminescence imaging; ROBO1 neutralizing antibody; human PDAC patient samples; mechanistic dependence receptor characterization |
Nature communications |
Medium |
36792623
|