Journal: Oncology Reports

Article Title: Berberine exerts its antineoplastic effects by reversing the Warburg effect via downregulation of the Akt/mTOR/GLUT1 signaling pathway

doi: 10.3892/or.2021.8204

Figure Lengend Snippet: Ubc9 and GIPC mediate the post-translational modification and cytoplasmic retention of GLUT1, respectively. MCF7 cells were pretreated with the (A) proteasome inhibitor MG-132 or (B) lysosomal inhibitor, leupeptin, and the expression levels of GLUT1 were analyzed using western blotting. Flag-tagged GLUT1, His-tagged Ubc9 and His-tagged GIPC were constructed, and the interactions between (C) Ubc9 and GLUT1 and (D) GIPC and GLUT1 were analyzed using co-immunoprecipitation. Data are presented as the mean ± SD; n=3. **P<0.01 and ## P<0.01. Ubc9, ubiquitin conjugating enzyme E2 I; GIPC, Gα-interacting protein-interacting protein at the C-terminus; GLUT1, glucose transporter 1; Con, control; His, histidine; WCL, whole cell lysate.

Article Snippet: The open reading frames (ORFs) of GLUT1, ubiquitin conjugating enzyme E2 I (Ubc9) and Gα-interacting protein-interacting protein at the C-terminus (GIPC) were amplified with PrimerSTAR ® Max DNA Polymerase (cat. no. R045A; Takara Bio, Inc.) using cDNA as a template.

Techniques: Modification, Expressing, Western Blot, Construct, Immunoprecipitation

Journal: Oncology Reports

Article Title: Berberine exerts its antineoplastic effects by reversing the Warburg effect via downregulation of the Akt/mTOR/GLUT1 signaling pathway

doi: 10.3892/or.2021.8204

Figure Lengend Snippet: Proposed underlying mechanism of the antineoplastic effects of BBR, which involves the reversal of the Warburg effect via downregulating the Akt/mTOR/GLUT1 signaling pathway. Treatment with BBR downregulates the expression levels of p-Akt in cancer cells, which in turn downregulates the levels of its downstream signaling protein, p-mTOR. Subsequently, the binding between GIPC and GLUT1 is weakened, which results in the cytoplasmic retention of GLUT1. The weakened binding of GIPC with GLUT1 also strengthens the binding between Ubc9 and GLUT1, which leads to the post-translational degradation of GLUT1 and further diminishes the glucose transport function of GLUT1. Consequently, the glucose uptake capacity of cancer cells and ATP synthesis are decreased, therefore the Warburg effect of cancer cells is reversed, which contributes to the antineoplastic activity of BBR. BBR, berberine; Ubc9, ubiquitin conjugating enzyme E2 I; GIPC, Gα-interacting protein-interacting protein at the C-terminus; GLUT1, glucose transporter 1; p-, phosphorylated.

Article Snippet: The open reading frames (ORFs) of GLUT1, ubiquitin conjugating enzyme E2 I (Ubc9) and Gα-interacting protein-interacting protein at the C-terminus (GIPC) were amplified with PrimerSTAR ® Max DNA Polymerase (cat. no. R045A; Takara Bio, Inc.) using cDNA as a template.

Techniques: Expressing, Binding Assay, Activity Assay

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: iScience

Article Title: Spatial-temporal regulation of the prostanoid receptor EP2 co-ordinates PGE2-mediated cAMP signaling in decidualizing human endometrium

doi: 10.1016/j.isci.2024.111170

Figure Lengend Snippet:

Article Snippet: GIPC: sense 5′-GTCCAGACAGCAGCCAGAAT-3′ GIPC: antisense 5′-GAGGCCCTGTGATGTGAAGT-3′ , Merck , KSPQ12012G.

Techniques: Recombinant, Bioassay, Sequencing, Expressing, RNA Sequencing Assay, Cell Culture, Software, esiRNA, Negative 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) 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: