Journal: eLife

Article Title: GIPC proteins negatively modulate Plexind1 signaling during vascular development

doi: 10.7554/eLife.30454

Figure Lengend Snippet: ( A ) Confocal lateral images of the trunk vasculature (green) of 32 hpf embryos (region dorsal to the yolk extension). Anterior, left; dorsal, up. Scale bars (white horizontal lines), 100 μm. Genotypes indicated on top of each image in yellow font. Angiogenesis deficits are indicated as follows: white asterisks (DLAV gaps), magenta asterisks (truncated Se), white greater-than sign (thin Se). Maternal-zygotic (MZ) removal of gipc activity is denoted by the designation ‘MZ’ in superscript. In the WT image (top left), the vessels are designated with the white font as follows: DLAV (Dorsal Longitudinal Anastomotic Vessel), Se (Segmental Vessel), DA (Dorsal Aorta), and PCV (Posterior Cardinal Vein). ( B ) Bar graph. Percentage of Se in 32 hpf embryos of the indicated genotypes belonging to each of the following four phenotypic classes. Truncated: severe (includes missing Se), medium (yellow), and weak (gray). Non-truncated: complete (black). Significance values were calculated using a two-sided Fisher Exact test and significant differences (p<0.0033) assigned using a Bonferroni-type adjustment for 15 pairwise genotype comparisons (0.05/15 = 0.0033). Brackets and asterisks indicate pairs of genotypes with significantly different distributions of these four phenotypic classes. Quantifications. We scored Se angiogenesis in embryos of the following six genotypes: WT (138 Se, 12 embryos; an average of 11.5 Se/embryo), gipc1 skt1 (130 Se, 11 embryos; an average of 11.8 Se/embryo) , gipc1 skt1(MZ) (380 Se, 33 embryos; an average of 11.5 Se/embryo) , gipc2 skt3/skt4 (130 Se, 11 embryos; an average of 11.8 Se/embryo) , gipc1 skt1 ; gipc2 skt3/skt4 (152 Se, 13 embryos; an average of 11.6 Se/embryo), and gipc1 skt1(MZ) ; gipc2 skt3/skt4 (220 Se, 19 embryos; an average of 11.5 Se/embryo). For additional data, graphs, and statistical comparisons related to this figure, see , and . Please note that given the use of different scales for scoring angiogenesis deficits, it is unfeasible to compare the quantifications in and directly.

Article Snippet: Antibody , Rabbit anti-GIPC1 , Proteintech Group , Cat. #14822–1-AP. RRID: AB_2263269 , WB (1:3,000). This antibody detects GIPC1, GIPC2, and GIPC3 (our data).

Techniques: Activity Assay

Journal: eLife

Article Title: GIPC proteins negatively modulate Plexind1 signaling during vascular development

doi: 10.7554/eLife.30454

Figure Lengend Snippet: ( A–D ) Confocal lateral images of the trunk vasculature (green) of 32 hpf embryos (region dorsal to the yolk extension). Anterior, left; dorsal, up. Scale bars (white horizontal lines), 100 μm. Morpholino injection (un-injected or injected with plxnd1 morpholino) indicated on top, genotypes (WT or gipc1 skt1(MZ) ; gipc2 skt4(MZ) ) indicated on the left. The un-injected WT picture ( A ) shows the names of the major vessels in white font: DLAV (Dorsal Longitudinal Anastomotic Vessel), Se (Segmental Vessel), DA (Dorsal Aorta), and PCV (Posterior Cardinal Vein). Vascular defects highlighted as follows: truncated or missing Se (magenta asterisk), thin Se (white greater/less-than signs), DLAV gaps (white asterisk). Quantifications. The following number of embryos were analyzed: WT (four embryos), WT injected with plxnd1 morpholino (four embryos; 4/4 showed a vascular phenotype similar to that of plxnd1 fov01b nulls), gipc1 skt1(MZ) ; gipc2 skt4(MZ) (12 embryos; 7/12 showed angiogenesis deficits), and gipc1 skt1(MZ) ; gipc2 skt4(MZ) injected with plxnd1 morpholino (11 embryos; 11/11 showed a vascular phenotype similar to that of plxnd1 fov01b nulls).

Article Snippet: Antibody , Rabbit anti-GIPC1 , Proteintech Group , Cat. #14822–1-AP. RRID: AB_2263269 , WB (1:3,000). This antibody detects GIPC1, GIPC2, and GIPC3 (our data).

Techniques: Injection

Journal: eLife

Article Title: GIPC proteins negatively modulate Plexind1 signaling during vascular development

doi: 10.7554/eLife.30454

Figure Lengend Snippet: ( A–F ). Representative fluorescent images of HUVEC morphology in cell collapse experiments under the following conditions. No ligand ( A–C ; top) or 45 min stimulation with 10 nM of SEMA3E ( D–F ; bottom). In each picture, the square area marked by yellow dotted sides is shown at twice the magnification at the bottom left corner and delimited by white sides. shRNA treatments as follows. Non-targeting, control ( A, D ), GIPC ( GIPC1, GIPC2, and GIPC3 ; B, E ), and PLXND1 ( C, F ). Scale bars (white horizontal lines), 100 μm. ( A–C ) Without ligand stimulation cells are uncollapsed regardless of the knockdown condition. ( D, E ) Cell collapse under ligand stimulation. Cells treated with non-targeting, control shRNA collapse ( D ). GIPC knockdown cells hypercollapse ( E ). SEMA3E-induced collapse is PLXND1-dependent. PLXND1 knockdown abrogates the morphological response ( F ). Cell collapse data collected from three independent experiments. This figure is related to .

Article Snippet: Antibody , Rabbit anti-GIPC1 , Proteintech Group , Cat. #14822–1-AP. RRID: AB_2263269 , WB (1:3,000). This antibody detects GIPC1, GIPC2, and GIPC3 (our data).

Techniques: shRNA, Control, Knockdown

Journal: eLife

Article Title: GIPC proteins negatively modulate Plexind1 signaling during vascular development

doi: 10.7554/eLife.30454

Figure Lengend Snippet: Western blots for GIPC1, GIPC2, PLXND1, and GAPDH (loading control) from TCLs of cells infected with the indicated shRNA lentiviral particles. Note the effective decrease of GIPC1-2 and PLXND1 levels. GIPC3 expression was absent under all the experimental conditions assayed. Hence, for brevity, the corresponding Western blots are not shown. This figure is related to .

Article Snippet: Antibody , Rabbit anti-GIPC1 , Proteintech Group , Cat. #14822–1-AP. RRID: AB_2263269 , WB (1:3,000). This antibody detects GIPC1, GIPC2, and GIPC3 (our data).

Techniques: Western Blot, Control, Infection, shRNA, Expressing

Journal: eLife

Article Title: GIPC proteins negatively modulate Plexind1 signaling during vascular development

doi: 10.7554/eLife.30454

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit anti-GIPC1 , Proteintech Group , Cat. #14822–1-AP. RRID: AB_2263269 , WB (1:3,000). This antibody detects GIPC1, GIPC2, and GIPC3 (our data).

Techniques: Transgenic Assay, Plasmid Preparation, Mutagenesis, Derivative Assay, Selection, Stable Transfection, Knock-Out, Recombinant, Control, shRNA, Sequencing, Construct, Synthesized, Positive Control, Sterility, Concentration Assay

Journal: eLife

Article Title: GIPC proteins negatively modulate Plexind1 signaling during vascular development

doi: 10.7554/eLife.30454

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit anti-GIPC1 , Proteintech Group , Cat. #14822–1-AP. RRID: AB_2263269 , WB (1:3,000). This antibody detects GIPC1, GIPC2, and GIPC3 (our data).

Techniques: Sequencing

Journal: medRxiv

Article Title: GIPC1 intermediate-length repeat expansion in amyotrophic lateral sclerosis

doi: 10.1101/2025.05.22.25328088

Figure Lengend Snippet: (A) The CGG repeat is located within exon 1 of GIPC1 . Primers and probes for repeat-primed PCR (RP-PCR, green), fragment analysis (blue), and RNA fluorescence in situ hybridization (FISH, red) were designed within or adjacent to the repeat region. (B) The left panel shows the results for RP-PCR, in which abnormal CGG repeat expansions were detected in the upper three cases. The middle panel displays the results of fragment analysis. The right panel presents histograms of repeat sizes estimated from long-read sequencing data using NanoRepeat; orange bars indicate expanded alleles. (C) Histogram of CGG repeat sizes determined through fragment analysis. The lower graph shows an enlarged view of the low-frequency range. In the control group, most alleles contained 32 or fewer repeats. Two control samples exhibited abnormally expanded alleles, whereas four ALS samples harbored alleles with 33 or more repeats.

Article Snippet: GIPC1 immunostaining of spinal cord sections from patients with ALS (A) Anterior horn of the lumbar spinal cord from an ALS patient with a CGG repeat expansion stained with anti-GIPC1 antibody (Proteintech). (B) Anterior horn of the lumbar spinal cord from an ALS patient without a CGG repeat expansion stained with anti-GIPC1 antibody (Proteintech). (C) Anterior horn of the lumbar spinal cord from an ALS patient with a CGG repeat expansion stained with anti-GIPC1 antibody (Santa Cruz Biotechnology). (D) Anterior horn of the lumbar spinal cord from an ALS patient without a CGG repeat expansion stained with anti-GIPC1 antibody (Santa Cruz Biotechnology).

Techniques: Fluorescence, In Situ Hybridization, Sequencing, Control

Journal: medRxiv

Article Title: GIPC1 intermediate-length repeat expansion in amyotrophic lateral sclerosis

doi: 10.1101/2025.05.22.25328088

Figure Lengend Snippet: (A) Results of RNA FISH using a (CGG) 8 probe. Intranuclear RNA foci were observed exclusively in the ALS patient harboring a CGG repeat expansion in GIPC1 . (B) RNA FISH results using another probe targeting the 5′ UTR of GIPC1 . Similar intranuclear RNA foci were detected with this GIPC1 -specific probe, confirming the presence of expanded repeat-containing RNA. Numbers in parentheses indicate the number of CGG repeats in GIPC1 . Scale bars: 5 μm.

Article Snippet: GIPC1 immunostaining of spinal cord sections from patients with ALS (A) Anterior horn of the lumbar spinal cord from an ALS patient with a CGG repeat expansion stained with anti-GIPC1 antibody (Proteintech). (B) Anterior horn of the lumbar spinal cord from an ALS patient without a CGG repeat expansion stained with anti-GIPC1 antibody (Proteintech). (C) Anterior horn of the lumbar spinal cord from an ALS patient with a CGG repeat expansion stained with anti-GIPC1 antibody (Santa Cruz Biotechnology). (D) Anterior horn of the lumbar spinal cord from an ALS patient without a CGG repeat expansion stained with anti-GIPC1 antibody (Santa Cruz Biotechnology).

Techniques:

Journal: Nature Communications

Article Title: Allosteric control of dynamin-related protein 1 through a disordered C-terminal Short Linear Motif

doi: 10.1038/s41467-023-44413-6

Figure Lengend Snippet: a CL-stimulated GTPase activities of WT and ΔCT6 Drp1 with increasing concentrations of GIPC-1. k cat from two independent measurements are plotted versus Drp1:GIPC-1 molar ratio. b Data in a replotted as % of maximum activity for each variant. c Representative NS-EM images of WT Drp1 and CT variants incubated with GMP-PCP in the absence and presence of a 1:4 molar ratio of GIPC-1. Scale bar, 100 nm. d NS-EM 2D class averages of WT Drp1 in the apo state in the presence of a 1:4 molar ratio of GIPC-1. Only the compact conformers (open and closed) were observed for WT-Drp1 in the presence of GIPC-1. e Kymograph showing NT constriction and fission (arrowhead) by WT Drp1 in the presence of GIPC-1 at a 1:1 molar ratio (0.5 μM each) in the presence of 1 mM GTP. NT membrane fluorescence is displayed in cyan pseudocolor for clarity. f Percentage of NTs that underwent fission upon addition of either 0.5 μM WT Drp1 alone or a 1:1 mixture of WT Drp1:GIPC-1 at 0.5 μM each in the presence of 1 mM GTP. Each point represents a replicate. The number on top of each column represents the total number of NTs for each condition. Mean ± SD are shown. ** Statistically different at the 0.01 level (unpaired two sample t -test, equal variance assumed). g Images showing the rigid polymerization of WT Drp1 alone versus the formation of much shorter scaffolds in the equimolar presence of WT Drp1 and GIPC-1 on NTs. Images shown were acquired approximately 2 minutes after the addition of the proteins at 2 μM final concentration each in the presence of 1 mM GTP. RhPE channel is shown. Pseudocolor is used for clarity.

Article Snippet: GST-tagged mouse GIPC-1 (sourced from Addgene (Plasmid #35791)) subcloned in pGEX-6P1 (Cytiva) was expressed and purified using standard protocols.

Techniques: Activity Assay, Variant Assay, Incubation, Membrane, Fluorescence, Concentration Assay

Journal: Molecular Pharmacology

Article Title: Modulation of ?-Opioid Receptor Signaling by RGS19 in SH-SY5Y Cells

doi: 10.1124/mol.112.081992

Figure Lengend Snippet: GIPC protein changes with RGS19. Whole-cell lysates were prepared from SH-SY5Y cells stably expressing either GFP shRNA or RGS19 shRNA as described under Materials and Methods. GIPC was detected as a single band at ∼37 KDa. This was decreased to ∼50% of control level in cells expressing shRNA against RGS19 compared with cells expressing shRNA against GFP (A). When SH-SY5Y cells were treated with (+) DAMGO (10 µM) overnight, GIPC was increased ∼50% compared with cells without DAMGO treatment (B). Quantified data from three sets of individual experiment are presented on the right side of the figure. Loading controls (α-tubulin) were not changed.

Article Snippet: Monoclonal anti-human GIPC antibody (clone AT1G10) was purchased from ATGen Co., Ltd. (Gyeonggi-do, South Korea).

Techniques: Stable Transfection, Expressing, shRNA, Control