Journal: Cancer Management and Research

Article Title: Inhibiting GIT1 reduces the growth, invasion, and angiogenesis of osteosarcoma

doi: 10.2147/CMAR.S181066

Figure Lengend Snippet: GIT1 expression in human osteosarcoma tissues. Notes: Two typical cases of primary osteosarcoma. In ( A ) there is a diffuse cytoplasmic staining, while no immunoreactivity can be detected in ( B ). No nuclear staining for GIT1 has been obtained (original magnification: ×400).

Article Snippet: The blots were blocked with 5% BSA, followed by incubation with antibodies against ERK1/2, p-ERK1/2, HIF-1α, GIT1, and GAPDH (Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C.

Techniques: Expressing, Staining

Journal: Cancer Management and Research

Article Title: Inhibiting GIT1 reduces the growth, invasion, and angiogenesis of osteosarcoma

doi: 10.2147/CMAR.S181066

Figure Lengend Snippet: Immunohistochemical staining of GIT1 in all patientderived OS tissues

Article Snippet: The blots were blocked with 5% BSA, followed by incubation with antibodies against ERK1/2, p-ERK1/2, HIF-1α, GIT1, and GAPDH (Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C.

Techniques: Immunohistochemical staining, Staining

Journal: Cancer Management and Research

Article Title: Inhibiting GIT1 reduces the growth, invasion, and angiogenesis of osteosarcoma

doi: 10.2147/CMAR.S181066

Figure Lengend Snippet: Knockdown efficacy of GIT1. Notes: ( A ) The relative transcription of GIT1 mRNA in three kinds of osteosarcoma cell lines (HOS, 143B and OST) detected by qRT-PCR. ( B ) The knockdown efficacy by GIT1-shRNA in OST cells detected by real-time RT-PCR. ( C ) The relative expression of GIT1 protein in three kinds of osteosarcoma cell lines (HOS, 143B and OST) detected by Western Blotting. ( D ) The knockdown efficacy by GIT1-shRNA in OST cells detected by Western Blotting. ( E ) Microscopic images of OST infected with lentivirus. One-way ANOVA for comparing GIT1 mRNA among HOS, 143B and OST cells or among blank, NC-shRNA and GIT1-shRNA groups was conducted respectively (scale bar: 100 µm; * P <0.05).

Article Snippet: The blots were blocked with 5% BSA, followed by incubation with antibodies against ERK1/2, p-ERK1/2, HIF-1α, GIT1, and GAPDH (Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C.

Techniques: Knockdown, Quantitative RT-PCR, shRNA, Expressing, Western Blot, Infection

Journal: Cancer Management and Research

Article Title: Inhibiting GIT1 reduces the growth, invasion, and angiogenesis of osteosarcoma

doi: 10.2147/CMAR.S181066

Figure Lengend Snippet: Knockdown of GIT1 inhibited invasion of osteosarcoma cells. Notes: OST cells were infected with GIT1-shRNA, NC-shRNA or blank control for 24 and 48 hours respectively. Cell invasion was evaluated by the transwell migration assay. ( A ) The representative images of three groups. ( B ) Quantitative results of cell invasion assay. One-way ANOVA for comparing the number of invade cells among blank, NC-shRNA and GIT1-shRNA group was conducted (scale bar: 20 µm; * P <0.05).

Article Snippet: The blots were blocked with 5% BSA, followed by incubation with antibodies against ERK1/2, p-ERK1/2, HIF-1α, GIT1, and GAPDH (Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C.

Techniques: Knockdown, Infection, shRNA, Control, Transwell Migration Assay, Invasion Assay

Journal: Cancer Management and Research

Article Title: Inhibiting GIT1 reduces the growth, invasion, and angiogenesis of osteosarcoma

doi: 10.2147/CMAR.S181066

Figure Lengend Snippet: Knockdown of GIT1 inhibited angiogenesis of osteosarcoma. Notes: ( A ) The ELISA results of VEGF concentration released from OST cells infected with GIT1-shRNA or NC-shRNA. ( B ) The representative images of immunohistochemistry of CD34 (a marker of tumor MVD) in vivo. ( C ) The statistical analysis of B. The independent Student’s t -test for comparing VEGF concentration or CD34 expression level between the two groups was conducted respectively (Scale bar: 20 µm; * P <0.05).

Article Snippet: The blots were blocked with 5% BSA, followed by incubation with antibodies against ERK1/2, p-ERK1/2, HIF-1α, GIT1, and GAPDH (Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C.

Techniques: Knockdown, Enzyme-linked Immunosorbent Assay, Concentration Assay, Infection, shRNA, Immunohistochemistry, Marker, In Vivo, Expressing

Journal: Cancer Management and Research

Article Title: Inhibiting GIT1 reduces the growth, invasion, and angiogenesis of osteosarcoma

doi: 10.2147/CMAR.S181066

Figure Lengend Snippet: Effect of GIT1 on primary osteosarcoma growth and lung metastasis in the orthotopic model. Notes: OST cells infected with GIT1-shRNA or NC-shRNA were orthotopically transplanted into the distal femur of nude mice and allowed to form tumors. ( A ) Tumor diameter was evaluated for 45 days. ( B ) The representative fluorescence imaging of orthotopic tumor in two groups. The independent Student’s t -test for comparing tumor volume between NC-shRNA and GIT1-shRNA group was conducted in different post-implantation days. ( C ) The representative fluorescence imaging of lung metastasis in two groups. Left column: the images of whole mice; right column: the images of lung metastasis (* P <0.05).

Article Snippet: The blots were blocked with 5% BSA, followed by incubation with antibodies against ERK1/2, p-ERK1/2, HIF-1α, GIT1, and GAPDH (Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C.

Techniques: Infection, shRNA, Fluorescence, Imaging

Journal: Cancer Management and Research

Article Title: Inhibiting GIT1 reduces the growth, invasion, and angiogenesis of osteosarcoma

doi: 10.2147/CMAR.S181066

Figure Lengend Snippet: Data of lung metastasis

Article Snippet: The blots were blocked with 5% BSA, followed by incubation with antibodies against ERK1/2, p-ERK1/2, HIF-1α, GIT1, and GAPDH (Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C.

Techniques:

Journal: Cancer Management and Research

Article Title: Inhibiting GIT1 reduces the growth, invasion, and angiogenesis of osteosarcoma

doi: 10.2147/CMAR.S181066

Figure Lengend Snippet: ERK1/2/HIF-1α pathway participated in GIT1-regulated VEGF release. Notes: ( A ) The activation of ERK1/2 in tumor tissues from two groups’ mice. ( B ) The statistical analysis of A. ( C ) The expression of HIF-1α in tumor tissues from two groups` mice. ( D ) The statistical analysis of C. ( E ) In order to induce HIF-1α expression, OST cells of two groups were cultured in hypoxia for 12 hours with or without PD98059 pretreated. The expression of HIF-1α under different conditions was detected by Western blotting. ( F ) The statistical analysis of C. ( G ) The ELISA results of VEGF concentration released from OST cells infected with GIT1-shRNA with or without PD98059 pretreated. The independent Student’s t -test for comparing p-EKR1/2 or HIF-1α protein level between NC-shRNA and GIT1-shRNA group was conducted, respectively (* P <0.05).

Article Snippet: The blots were blocked with 5% BSA, followed by incubation with antibodies against ERK1/2, p-ERK1/2, HIF-1α, GIT1, and GAPDH (Cell Signaling Technology, Danvers, MA, USA) overnight at 4°C.

Techniques: Activation Assay, Expressing, Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay, Concentration Assay, Infection, shRNA

Journal: Neural Regeneration Research

Article Title: GIT1 enhances neurite outgrowth by stimulating microtubule assembly

doi: 10.4103/1673-5374.179054

Figure Lengend Snippet: GIT1 was highly expressed in the nervous system. (A) Expression profiles of Git1 and Git2 in 2-month-old mice, as assessed by RT-PCR. Ctrl: Negative control. (B) The relative mRNA expression levels of Git1 and Git2 in 2-month-old mice, normalized to Gapdh . The data are expressed as optical density ratio of Git / Gapdh ( n = 3; data are given as mean ± SEM). (C) GIT1 protein expression in the whole brain from embryonic day (E) 10.5 to postnatal day (P) 60. (D) GIT1 expression in the mouse hippocampus at P0, 7, 14, 30 and 60. Quantification of GIT1 expression in the whole brain (E) and hippocampus (F) were normalized to β-actin. Data represent the mean ± SEM of three independent observations.

Article Snippet: Immediately, 1 or 5 μM GIT1 protein (Proteintech) diluted in tubulin buffer was added to the wells.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Negative Control

Journal: Neural Regeneration Research

Article Title: GIT1 enhances neurite outgrowth by stimulating microtubule assembly

doi: 10.4103/1673-5374.179054

Figure Lengend Snippet: Neurite growth was impaired in GIT1 knockout neurons. (A) Hippocampal neurons isolated from wild type (WT), GIT1 heterozygote (HET) and GIT1 knockout (KO) mice at postnatal day (P) 0 were cultured for 5 days in control medium (left) or medium containing 50 ng/mL nerve growth factor (NGF) (right). Neurons were immunostained with anti-Tuj1 (green) antibody and DAPI (blue). Scale bar: 20 μm. Quantification of total neurite length per neuron (B) and average length of axon-like processes (D) of WT, HET and KO hippocampal neurons in vitro . The effects of NGF treatment are shown in Figure D. n = 30–50 cells; data are expressed as the mean ± SEM. The experiments were repeated three times. (C) Western blot of lysates from P0 WT and HET hippocampal neurons cultured for 5 days in the presence of 0, 25 or 50 ng/mL NGF. Anti-GIT1 and anti-actin antibodies were used to detect target protein expression. * P < 0.05, ** P < 0.01, *** P < 0.001. One-way analysis of variance followed by Tukey's test.

Article Snippet: Immediately, 1 or 5 μM GIT1 protein (Proteintech) diluted in tubulin buffer was added to the wells.

Techniques: Knock-Out, Isolation, Cell Culture, Control, In Vitro, Western Blot, Expressing

Journal: Neural Regeneration Research

Article Title: GIT1 enhances neurite outgrowth by stimulating microtubule assembly

doi: 10.4103/1673-5374.179054

Figure Lengend Snippet: Axonal growth in GIT1 knockout (KO) hippocampal neurons and the effects of overexpression of GIT1 or GIT1 deletion constructs. (A) Schematic diagram of the full-length GIT1 and different GIT1 deletion constructs: cGIT1, CDΔAS/SLD, nGIT1 and GIT1ΔSLD. The Flag-tag was attached to the N terminal of the constructs. (B) KO hippocampal neurons were cultured for 5 days after transfection with the constructs, and then stained with anti-Flag (green) and anti-Tuj1 (red) antibodies and DAPI (blue). Scale bars: 20 μm. (C) Average length of axon-like processes in KO hippocampal neurons cultured for 5 days after transfection with the GIT1 deletion constructs. Cells were analyzed using ImageJ software. n = 3 cells; data are expressed as the mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001. One-way analysis of variance followed by Tukey's test. The experiments were repeated three times. Con: Control.

Article Snippet: Immediately, 1 or 5 μM GIT1 protein (Proteintech) diluted in tubulin buffer was added to the wells.

Techniques: Knock-Out, Over Expression, Construct, FLAG-tag, Cell Culture, Transfection, Staining, Software, Control

Journal: Neural Regeneration Research

Article Title: GIT1 enhances neurite outgrowth by stimulating microtubule assembly

doi: 10.4103/1673-5374.179054

Figure Lengend Snippet: GIT1 interacted with tubulins and microtubule-associated proteins, and stimulated microtubule assembly. (A) Brain extract from 8-week-old mice was immunoprecipitated with the GIT1 antibody, and western blots were probed by antibodies against CRMP2, Clasp2, α-tubulin, Tuj1, TUBGCP3, MAP2, Tau, MAP1B and GIT1. The images show the colocalization of Tuj1 (red) and GIT1 (green) (B), Clasp2 (red) and GIT1 (green) (C), and MAP2 (red) and GIT1 (green) (D) in primary cultured hippocampal neurons on postnatal day (P) 0. Scale bars: 20 μm. (E) Expression levels of Tuj1, MAP2 and GIT1 in the cortex and hippocampus of wild-type (WT) and GIT1 heterozygote (HET) mice on postnatal day (P) 0, P15 and P30. (F) GIT1 activates tubulin polymerization in vitro . Result represents means of three independent observations. min: Minutes.

Article Snippet: Immediately, 1 or 5 μM GIT1 protein (Proteintech) diluted in tubulin buffer was added to the wells.

Techniques: Immunoprecipitation, Western Blot, Cell Culture, Expressing, In Vitro

Journal: bioRxiv

Article Title: JNK regulates GABAAR expression at the cell surface via the receptor clustering protein GIT1 (ArfGAP1)

doi: 10.1101/2025.01.22.634236

Figure Lengend Snippet: A. In vitro phosphorylation of brain homogenate by recombinant active JNK1 revealed phosphorylation of GIT1 on S371 and S692. Mass spectrometry-identified sequences are shown for GIT1 (accession NP_001078923.1). Β. Representative autoradiograph and Coomassie Brilliant Blue (CBB) stained gel image are shown for recombinant GST-GIT1- wild-type (WT), -S371A, -S692A or S371A/S692A (SAA) variants after phosphorylation by GST-JNK1. JNK1 phosphorylated GIT1-WT and GIT1–S692A but not GIT1-S371A, indicating that S371 is the preferred JNK1 phosphorylation site. Phosphorylation of GIT1 by JNK1 was prevented by JNK inhibitor 10 µM SP600125. C. Quantitative data from 4 repeats of experiment shown in B . D. Domain map of human GIT1 (UniProt identifier: Q9Y2X7-3) shows predicted JNK1 phosphorylation sites (yellow circles). Abbreviations: Arf GTPase activating protein domain (GAP), 3x Ankyrin repeats (ANK), Spa-Homology domain (SHD), Synapse Localization Domain (SLD) and Paxillin Binding Domain (PBD). E. Quantitative LC-MS/MS data of GIT1-S371 phosphorylation in wild-type (WT) and Jnk1-/- whole brains. F. Quantitative LC-MS/MS data of GIT1-S371 phosphorylation in brains from DJNKI-1- infused adult mice. G. Active Arf1 was detected from pull downs using GST-GAT-GGA3 immobilized on glutathione beads. Cells expressed GIT1-variants together with Arf1-WT or constitutively active Arf1-Q71L. H. Quantitative data from 3 repeats of experiment described in G. Phosphorylation-site mutants of GIT1 did not alter Arf1 activity. Error bars represent standard error of the mean (SEM) and P-values were determined using Student’s t test.

Article Snippet: pEGFP-C1 GIT1 (EGFP GIT1 WT) (Addgene plasmid: # 15226) and Flag-beta PIXa (Addgene plasmid: # 15234) were gifts from Rick Horwitz and pSuper-GIT1 shRNA was from Huaye Zhang.

Techniques: In Vitro, Recombinant, Mass Spectrometry, Autoradiography, Staining, Binding Assay, Liquid Chromatography with Mass Spectroscopy, Activity Assay

Journal: bioRxiv

Article Title: JNK regulates GABAAR expression at the cell surface via the receptor clustering protein GIT1 (ArfGAP1)

doi: 10.1101/2025.01.22.634236

Figure Lengend Snippet: A. Representative maximum projection images (25x optical slices, z=0.173 µm) depict EGFP-GIT1-WT and mRub-Lifeact in 16-DIV hippocampal neurons. Regions of interest (R.O.I.s; circles) depict where in the dendrite, spine and background mean intensity signals were acquired. B, D. Relative enrichment of EGFP-GIT1 variants in spines or dendrites, was calculated as a ratio of the mean intensity from the spine R.O.I. to the mean intensity from the R.O.I in the dendrite, after background subtraction, for the given treatments Jnk1-/- or TAT/DJNKI-1 (20 µM), 8 hours. C, E. Relative enrichment of mRuby-Lifeact fluorescence in spines was measured as for B and E. Mean data +/- S.E.Ms from a total of ∼80 spines from several neurons per condition are shown. Neurons originated from at least two independent mouse litters. P-values were calculated from Student’s two-tailed t test. F, G. We next checked the overall effect of DJNKI-1 inhibitor or Jnk1 knockout on GABAAR expression at the cell surface in the dendritic compartment. Both conditions increased general surface expression of GABAAR. H. In order to test the requirement for GIT1 in GABAAR trafficking, we characterised shRNA targeting GIT1. I. Representative maximum projection images of GABAAR β3 staining from 25 x optical sections (z=0.173 μm) are shown for WT and Jnk1-/- hippocampal neurons at 16 DIV. Staining was performed in non-permeabilized cells expressing mRuby, with non-targeting (NT)-shRNA or GIT1-shRNA and EGFP-GIT1 variants, as shown. White outlines were traced from mRuby fluorescence. Surface staining of GABAAR β3 is shown in pseudocolour. Representative images of total GABAAR β3 staining from an independent set of permeabilized cells is shown for comparison. J, K . Quantitative data from these experiments is shown for spines and dendrites. Mean data +/- S.E.M are shown from ∼70 dendritic spines per condition, in cells from at least two independent mouse litters. L, M. Cell surface expression of GABAAR β3 was measured from biotinylated samples. Representative blots show GABAAR β3 of lysates from before streptavidin-biotin enrichment (total) or after streptavidin-biotin enrichment (surface) are shown. Mean data from 3 repeats +/- S.E.M are shown. P-values were calculated from Student’s two-tailed t test.

Article Snippet: pEGFP-C1 GIT1 (EGFP GIT1 WT) (Addgene plasmid: # 15226) and Flag-beta PIXa (Addgene plasmid: # 15234) were gifts from Rick Horwitz and pSuper-GIT1 shRNA was from Huaye Zhang.

Techniques: Fluorescence, Two Tailed Test, Knock-Out, Expressing, shRNA, Staining, Comparison

Journal: bioRxiv

Article Title: JNK regulates GABAAR expression at the cell surface via the receptor clustering protein GIT1 (ArfGAP1)

doi: 10.1101/2025.01.22.634236

Figure Lengend Snippet: A. Representative maximum projection images (32 x optical sections of 0.173 μm) of neurons expressing mRuby to visualize morphology. Neurons at 16 DIV were immunostained for endogenous GIT1 (green) and paxillin (PXN), 14-3-3ζ or β-PIX (magenta). B. Quantitative data show mean intensities of GIT1, PXN, 14-3-3ζ and β-PIX in spines. DJNKI- 1 increases GIT1 in dendritic spines compared to control (TAT), and reduces β-PIX in the same cell compartment. C. Mean intensity ratios of GIT1, PXN, 14-3-3ζ and β-PIX in dendritic spine/shaft. Mean data +/- SEMs from ∼42 spines per condition.

Article Snippet: pEGFP-C1 GIT1 (EGFP GIT1 WT) (Addgene plasmid: # 15226) and Flag-beta PIXa (Addgene plasmid: # 15234) were gifts from Rick Horwitz and pSuper-GIT1 shRNA was from Huaye Zhang.

Techniques: Expressing, Control

Journal: bioRxiv

Article Title: JNK regulates GABAAR expression at the cell surface via the receptor clustering protein GIT1 (ArfGAP1)

doi: 10.1101/2025.01.22.634236

Figure Lengend Snippet: A. We examined whether GABAAR β3, paxillin, β-PIX or 14-3-3ζ co-immunoprecipitated with GIT1 using cortex homogenates from WT and Jnk1-/- adult mice. +/- denotes addition or not of antibody. Representative immunoblots of input and GIT1 immunoprecipitates probed with indicated antibodies are shown. B. Quantitative data show the extent of GABAAR β3, PXN, β-PIX and 14-3-3ζ co-purification with GIT1 from brain. For 14-3-3ζ quantification, the background bands present in samples without GIT1 antibody were subtracted from bands in samples with GIT1 antibody. There was increased interaction between GIT1 and GABAAR β3, PXN and 14-3-3ζ in Jnk1-/- brain compared to WT. In contrast, interaction between GIT1 and β-PIX decreased. Histogram bars represent mean data from 4 experimental repeats +/- SEM. C. The effect of TAT or DJNKI-1 on GIT1 interaction with binding partners was determined. Cortical neurons at 16 DIV were treated with 20 µM TAT or DJNKI-1 for 8 hours. Representative immunoblots of GIT1 co-precipitations are shown. D. Band quantification from blots in . DJNKI-1-treated neurons show increased interaction between GIT1 and GABAAR β3, paxillin and 14-3-3ζ, whereas β-PIX interaction with GIT1 reduced. Bars show mean data from 5 repeats ± SEM. E. We tested whether GIT1-S371 phosphorylation site mutants altered these interactions. HEK-293 cells were transfected with EGFP-GIT1-WT, EGFP-GIT1-S371A or EGFP-GIT1-S371D and either Flag (as a control), Flag-PXN-WT or Flag-PXN-S178A, as indicated. Representative blots of input and co-immunoprecipitations are shown. F. Quantitative band intensities from E are shown using log10 scale. PXN-S178A interacted most highly with GIT1-S371A. Mean data ± SEM from 4 repeats are shown. Student’s t-test p-values above bars are compared to samples with EGFP-GTI1-WT and Flag-PXN-WT. G. The same approach was used to test endogenous 14-3-3ζ and Flag-β-PIX interaction with GIT1. Representative blots are shown. H. Quantitative data for Flag-β−PIX interaction with EGFP-GIT1 variants. I. Quantitative data for 14-3-3ζ interaction with EGFP-GIT1 variants. Endogenous 14-3-3ζ interacts most highly with EGFP-GIT1-S371A. Bars ± SEM represent means from 3 repeats. p-values were calculated from Student’s two-tailed t-test and are compared to conditions where WT is expressed. IB: immunoblot, IP: immunoprecipitation.

Article Snippet: pEGFP-C1 GIT1 (EGFP GIT1 WT) (Addgene plasmid: # 15226) and Flag-beta PIXa (Addgene plasmid: # 15234) were gifts from Rick Horwitz and pSuper-GIT1 shRNA was from Huaye Zhang.

Techniques: Immunoprecipitation, Western Blot, Copurification, Binding Assay, Transfection, Control, Two Tailed Test

Journal: bioRxiv

Article Title: JNK regulates GABAAR expression at the cell surface via the receptor clustering protein GIT1 (ArfGAP1)

doi: 10.1101/2025.01.22.634236

Figure Lengend Snippet: A. Representative maximum projection images (22 x optical sections of 0.173 μm) showing staining of GABAAR β3 in non-permeabilised 16-DIV hippocampal neurons (magenta). Cells co-express mRuby with NT-shRNA or GIT1shRNA, as indicated. Cell tracing was based on mRuby images. Cells were treated with 10 µM R18 for 24 h and 20 µM TAT or DJNKI-1 for 8 h. B. Quantification of GABAAR β3 enrichment in dendritic spines. R18 prevented DJNKI-1-induced GABAAR β3 increase at the dendritic spines. C. R18 prevented GIT1-S371A-induced increase in GABAAR β3 at the cell surface. Histogram bars represent means from ∼51 spines per condition from several neurons. P-values were calculated from Student’s two-tailed t-tests were compared to control (TAT) unless otherwise indicated. D. Neurons were co-transfected at 7 DIV with EGFP-Dynamin 2-WT or EGFP-Dynamin 2- K44A and treated with 20 µM TAT or DJNKI-1 for 8 h. Representative images are shown. E. Mean intensities for GABAAR β3 are shown from non-permeabilised cells in mushroom spines. GABAAR surface expression increased in DJNKI-1-treated neurons, even when the K44A mutant is expressed. Mean data +/- SEMs from ∼70 spines per condition are shown. p- values (calculated from Student’s two-tailed t test) were compared to control (TAT) unless otherwise indicated by brackets.

Article Snippet: pEGFP-C1 GIT1 (EGFP GIT1 WT) (Addgene plasmid: # 15226) and Flag-beta PIXa (Addgene plasmid: # 15234) were gifts from Rick Horwitz and pSuper-GIT1 shRNA was from Huaye Zhang.

Techniques: Staining, shRNA, Two Tailed Test, Control, Transfection, Expressing, Mutagenesis

Journal: bioRxiv

Article Title: JNK regulates GABAAR expression at the cell surface via the receptor clustering protein GIT1 (ArfGAP1)

doi: 10.1101/2025.01.22.634236

Figure Lengend Snippet: We propose that when JNK1 active it phosphorylates GIT1 on S371. This reduces levels of GABAAR at the cell surface, both at excitatory synapses and at extrasynapatic sites. Conversely when JNK is inhibited or Jnk1 genetically deleted, GIT1 associates with a paxillin and 14-33-containing receptor complex at the membrane. GIT1 phosphorylation by JNK on S371 dissociates it from binding to this complex, leading to run-down of receptor surface expression. The consequence for neuronal physiology is increased inhibitor post synaptic currents and increased tonic inhibitory current.

Article Snippet: pEGFP-C1 GIT1 (EGFP GIT1 WT) (Addgene plasmid: # 15226) and Flag-beta PIXa (Addgene plasmid: # 15234) were gifts from Rick Horwitz and pSuper-GIT1 shRNA was from Huaye Zhang.

Techniques: Membrane, Binding Assay, Expressing