gapdh polyclonal antibody  (Proteintech)

Journal: iScience

Article Title: IPO5 contributes to cetuximab resistance in colorectal cancer by modulating Ras signaling

doi: 10.1016/j.isci.2026.117714

Figure Lengend Snippet: IPO5 is upregulated in CRC cell lines with decreased sensitivity to cetuximab (A) Dose-response curves and IC 50 values of cetuximab in six CRC cell lines. (B) Western blot analysis of IPO5 and EGFR in indicated cell lines (upper) and densitometric quantification relative to GAPDH (lower). (C) IPO5 expression in cetuximab-sensitive versus resistant CRC patients from GEO: GSE302922 . (D and E) Kaplan-Meier analysis of overall survival (D; GEO: GSE17538 ) and progression-free survival (E; GEO: GSE41258 ) stratified by IPO5 levels. Data in (A) and (B) are mean ± SD ( n = 3 independent experiments); data in (C) are presented as boxplots. Statistical significance was determined using non-linear regression (A), one-way ANOVA with Tukey’s test (B), unpaired two-tailed Student’s t test (C), or log rank test (D and E). Exact p values and hazard ratios (HRs) are indicated.

Article Snippet: GAPDH polyclonal antibody , Proteintech , Cat# 10494-1-AP; RRID: AB_2263076.

Techniques: Sensitivity, Western Blot, Analysis, Expressing, Two Tailed Test

Journal: iScience

Article Title: IPO5 contributes to cetuximab resistance in colorectal cancer by modulating Ras signaling

doi: 10.1016/j.isci.2026.117714

Figure Lengend Snippet: Knocking down IPO5 expression increases the sensitivity of colorectal cancer cells to cetuximab (A and B) Validation of IPO5 knockdown by siRNA/shRNA in RKO and SW620 cells (A) and adenovirus-mediated overexpression in Caco2 and LoVo cells (B) by western blotting. Densitometric quantification normalized to GAPDH is shown below each blot. (C and D) CCK-8 assay assessing cell viability after cetuximab treatment following IPO5 knockdown (C) or overexpression (D). (E and F) A colony formation assay was performed to evaluate proliferation in response to cetuximab treatment following IPO5 knockdown (E) or overexpression (F). Data are presented as mean ± SD ( n = 3). Statistical significance was determined using an unpaired two-tailed Student’s t test (shRNA in A and B), one-way ANOVA with Dunnett’s test (siRNA in A), or two-way ANOVA with Sidak’s test (C–F). Exact p values are indicated.

Article Snippet: GAPDH polyclonal antibody , Proteintech , Cat# 10494-1-AP; RRID: AB_2263076.

Techniques: Expressing, Sensitivity, Biomarker Discovery, Knockdown, shRNA, Over Expression, Western Blot, CCK-8 Assay, Colony Assay, Two Tailed Test

Journal: iScience

Article Title: IPO5 contributes to cetuximab resistance in colorectal cancer by modulating Ras signaling

doi: 10.1016/j.isci.2026.117714

Figure Lengend Snippet: Knocking down IPO5 increases cetuximab-induced apoptosis in colorectal cancer cells (A and B) Flow cytometry (annexin V/propidium iodide [PI]) analysis of cells after si-IPO5 transfection or IPO5 adenovirus transduction, followed by treatment with PBS or cetuximab (48 h). The quantification of apoptotic cells is shown. (C) Western blot analysis and densitometric quantification of active caspase-3, Bcl-2, and BAX levels following IPO5 modulation and cetuximab treatment. GAPDH served as a loading control. Data are presented as mean ± SD ( n = 3). Statistical significance was determined using two-way ANOVA with Sidak’s test (A–C). Exact p values are indicated.

Article Snippet: GAPDH polyclonal antibody , Proteintech , Cat# 10494-1-AP; RRID: AB_2263076.

Techniques: Flow Cytometry, Analysis, Transfection, Transduction, Western Blot, Control

Journal: iScience

Article Title: IPO5 contributes to cetuximab resistance in colorectal cancer by modulating Ras signaling

doi: 10.1016/j.isci.2026.117714

Figure Lengend Snippet: The knockdown of IPO5 reverses cetuximab resistance in colorectal cancer cells (A) Cell morphology (left) and dose-response curves with IC 50 values (right) of parental LoVo and cetuximab-resistant LoVo-CR cells. Scale bars, 50 μm. (B) Non-permeabilized immunofluorescence staining of cell-surface EGFR (red) and nucleus (DAPI; blue) in LoVo and LoVo-CR cells. Scale bars, 20 μm. (C and D) Western blot analysis and densitometric quantification (normalized to GAPDH) of HER2 and c-MET (C), as well as IPO5, p-EGFR, EGFR, p-AKT, AKT, and HRAS (D) expression in LoVo and LoVo-CR cells. (E) Western blot validation of IPO5 knockdown by siRNA or shRNA in LoVo-CR cells. (F and G) CCK-8 (F) and colony formation (G) assays evaluating cell viability and proliferation under cetuximab treatment following IPO5 knockdown in LoVo-CR cells. Data are presented as mean ± SD ( n = 3 independent experiments). Statistical significance was evaluated using non-linear regression (A), an unpaired two-tailed Student’s t test (C and D; shRNA knockdown in E), one-way ANOVA with Dunnett’s test (siRNA in E), or two-way ANOVA with Dunnett’s (F) or Sidak’s (G) post hoc test. Exact p values are indicated.

Article Snippet: GAPDH polyclonal antibody , Proteintech , Cat# 10494-1-AP; RRID: AB_2263076.

Techniques: Knockdown, Morphology, Immunofluorescence, Staining, Western Blot, Analysis, Expressing, Biomarker Discovery, shRNA, CCK-8 Assay, Colony Assay, Two Tailed Test

Journal: iScience

Article Title: IPO5 contributes to cetuximab resistance in colorectal cancer by modulating Ras signaling

doi: 10.1016/j.isci.2026.117714

Figure Lengend Snippet: The overexpression of IPO5 attenuates the cetuximab-mediated inhibition of Ras signaling (A) Subcellular fractionation and densitometric quantification of RASAL2 in cytoplasmic (Cyt) and nuclear (Nuc) fractions of RKO and LoVo cells following IPO5 knockdown or overexpression (lamin B and GAPDH served as fraction controls). (B) GST-Raf1-RBD pull-down assay assessing active Ras-GTP levels in CRC cells upon IPO5 modulation. (C) Rescue GST pull-down assays evaluating Ras-GTP levels in Caco2 and LoVo cells co-transfected with IPO5 and wild-type (WT) or NLS-mutant (Mut) RASAL2. (D) Western blot analysis and quantification of downstream proteins (p-AKT, AKT, p-MEK, MEK1/2, p-ERK, and ERK1/2) in cetuximab-treated cells under IPO5 modulation. (E) CCK-8 cell viability curves of Caco2 and LoVo cells treated with cetuximab following co-transfection of IPO5 with RASAL2-NLS-WT or NLS-Mut. Data are presented as mean ± SD ( n = 3 independent experiments). Statistical significance was determined using an unpaired two-tailed Student’s t test (A and B), one-way ANOVA with Tukey’s test (C), or two-way ANOVA with Tukey’s test (D and E). Exact p values are indicated.

Article Snippet: GAPDH polyclonal antibody , Proteintech , Cat# 10494-1-AP; RRID: AB_2263076.

Techniques: Over Expression, Inhibition, Cell Fractionation, Knockdown, Pull Down Assay, GST Pulldown Assay, Transfection, Mutagenesis, Western Blot, Analysis, CCK-8 Assay, Cotransfection, Two Tailed Test

Journal: iScience

Article Title: IPO5 contributes to cetuximab resistance in colorectal cancer by modulating Ras signaling

doi: 10.1016/j.isci.2026.117714

Figure Lengend Snippet:

Article Snippet: GAPDH polyclonal antibody , Proteintech , Cat# 10494-1-AP; RRID: AB_2263076.

Techniques: Recombinant, Virus, Lysis, CCK-8 Assay, Protein Extraction, Activation Assay, Microarray, Gene Expression, shRNA, Sequencing, Control, Negative Control, Plasmid Preparation, Software

Journal: iScience

Article Title: Domain-autonomous MKLP1 modules divergently regulate mitotic fidelity and polyploidy in a holocentric system

doi: 10.1016/j.isci.2026.117149

Figure Lengend Snippet: The BmMKLP1 domains exhibit divergent nuclear and cytoplasmic localization (A) Subcellular localization of transiently expressed GFP-tagged domains (green) in BmN4 cells. Representative merged bright-field and GFP fluorescence images acquired at 10×, scale bars, 100 μm, BmN4 cells expressing the indicated constructs were imaged by differential interference contrast (DIC) microscopy (left) to show cell morphology and by fluorescence microscopy (right) to detect GFP-tagged proteins. All images were acquired with a 40× objective lens. Scale bars, 20 μm. (B) Western blot analysis of protein extracts from transfected BmN4 cells probed with an anti-GFP antibody (1:1,000, Proteintech, Wuhan, China), confirming the production of each GFP-tagged domain at its expected molecular weight. The “control” lane corresponds to the cycle 3 GFP-bleo fusion protein (∼39–40 KDa) expressed from the pIZT/V5-His vector. A representative blot from three independent experiments is shown. The PageRuler Prestained Protein Ladder (10–180 kDa; Thermo Fisher Scientific, Waltham, MA, USA) was used as a molecular weight marker.

Article Snippet: Scale bars, 20 μm. (B) Western blot analysis of protein extracts from transfected BmN4 cells probed with an anti-GFP antibody (1:1,000, Proteintech, Wuhan, China), confirming the production of each GFP-tagged domain at its expected molecular weight.

Techniques: Fluorescence, Expressing, Construct, Microscopy, Morphology, Western Blot, Analysis, Transfection, Molecular Weight, Control, Plasmid Preparation, Marker

Journal: iScience

Article Title: Domain-autonomous MKLP1 modules divergently regulate mitotic fidelity and polyploidy in a holocentric system

doi: 10.1016/j.isci.2026.117149

Figure Lengend Snippet: The head and stalk domains induce G2/M arrest, whereas the tail promotes polyploidization (A) Representative flow cytometry histograms showing DNA content profiles (top) and polyploidy distributions (bottom) for control cells and cells expressing GFP-tagged BmMKLP1—full-length, head, -stalk, or -tail. Cell cycle phases (G0/G1, S, and G2/M) were determined by model-based fitting of propidium iodide (PI) fluorescence intensity histograms. Polyploidy histograms depict cells with DNA content exceeding 4 N. (B) Quantification of cell cycle distribution under each condition. Bars represent the mean ± SEM percentage of cells in G0/G1, S, and G2/M phases from at least three independent experiments. Different lowercase letters indicate significant differences (one-way ANOVA followed by Tukey’s HSD, p < 0.05). (C) Percentage of polyploid cells under each condition. Bars represent mean ± SEM, with statistical significance assessed as in (B).

Article Snippet: Scale bars, 20 μm. (B) Western blot analysis of protein extracts from transfected BmN4 cells probed with an anti-GFP antibody (1:1,000, Proteintech, Wuhan, China), confirming the production of each GFP-tagged domain at its expected molecular weight.

Techniques: Flow Cytometry, Control, Expressing, Fluorescence

Journal: iScience

Article Title: Domain-autonomous MKLP1 modules divergently regulate mitotic fidelity and polyploidy in a holocentric system

doi: 10.1016/j.isci.2026.117149

Figure Lengend Snippet:

Article Snippet: Scale bars, 20 μm. (B) Western blot analysis of protein extracts from transfected BmN4 cells probed with an anti-GFP antibody (1:1,000, Proteintech, Wuhan, China), confirming the production of each GFP-tagged domain at its expected molecular weight.

Techniques: Transfection, Cloning, Imaging, Bicinchoninic Acid Protein Assay, Western Blot, Plasmid Preparation, Recombinant, Software

Journal: iScience

Article Title: M2 type tumor-associated macrophage-derived CCL-18 promotes glioma cell proliferation but does not mediates glioma recurrence after radiotherapy

doi: 10.1016/j.isci.2026.117695

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Article Snippet: Anti-p105/p50 rabbit antibody , Proteintech , 14220-1-AP; RRID: AB_2153393.

Techniques: Cell-Signaling, Virus, Tissue, Microarray, Recombinant, cDNA Synthesis, Enzyme-linked Immunosorbent Assay, Software, Biomarker Discovery, In Vivo Imaging

Journal: iScience

Article Title: Sulfonylurea receptor 1 deficiency alleviates acute lung injury by activating PARP1 in epithelial cells

doi: 10.1016/j.isci.2026.117627

Figure Lengend Snippet:

Article Snippet: Anti-PARP1 , Proteintech Group , Cat# 13371-1-AP; RRID: AB_2160459.

Techniques: Cell-Signaling, Recombinant, Transfection, Caspase-3 Activity Assay, Protein Extraction, Knock-Out, Sequencing, Control, Plasmid Preparation, Software, Analysis

Journal: iScience

Article Title: TXNRD1-driven enteric glial remodeling promotes neuronal dysfunction in Hirschsprung disease

doi: 10.1016/j.isci.2026.117636

Figure Lengend Snippet:

Article Snippet: Rabbit anti-SPP1 , Proteintech , Cat# 22952-1-AP; RRID: AB_2783651.

Techniques: Model Organisms, Recombinant, CCK-8 Assay, Apoptosis Assay, ROS Assay, Calcium Assay, Enzyme-linked Immunosorbent Assay, Generated, Software, Genotyping

Journal: iScience

Article Title: Domain-autonomous MKLP1 modules divergently regulate mitotic fidelity and polyploidy in a holocentric system

doi: 10.1016/j.isci.2026.117149

Figure Lengend Snippet:

Article Snippet: rabbit anti-β-tubulin , Proteintech, Wuhan, China , Cat#10094-1-AP;RRID:AB_2210695.

Techniques: Transfection, Cloning, Imaging, Bicinchoninic Acid Protein Assay, Western Blot, Plasmid Preparation, Recombinant, Software

Journal: iScience

Article Title: IPO5 contributes to cetuximab resistance in colorectal cancer by modulating Ras signaling

doi: 10.1016/j.isci.2026.117714

Figure Lengend Snippet:

Article Snippet: The primary antibodies used in this study were as follows: anti-IPO5 (1.0 μg/mL, Bioworld, China, #BS7341), anti-GAPDH (0.2 μg/mL, Proteintech, USA, #10494–1-AP), anti-AKT (1.0 μg/mL, Bioworld, China, #AP0489), anti- p -AKT (1.5 μg/mL, Bioworld, China, #BS4255), anti- p -EGFR (1.0 μg/mL, Bioworld, China, #BS6352), anti-active Caspase3 (1.5 μg/mL, Bioworld, China, #BS7004), anti-MEK1/2 (0.8 μg/mL, Affinity, USA, #AF6385), anti- p -MEK1/2 (1.0 μg/mL, Bioss, Beijing, #bsm-55741 R), anti-BCL-2 (0.5 μg/mL,Proteintech, USA, #12789-1-AP), anti-RASAL2 (1.2 μg/mL, Immunway, USA, #YN1852), anti- p -ERK (1.0 μg/mL, Immunway, USA, #YP0101), anti-ERK (0.8 μg/mL, Immunway, USA, #YT1625), anti-BAX (1.0 μg/mL, Immunway, USA, #YT0455), anti-EGFR (1.0 μg/mL, Abcam, USA, #ab52894), and anti-HRAS (0.5 μg/mL, Abcam, USA, #ab191595).

Techniques: Recombinant, Virus, Lysis, CCK-8 Assay, Protein Extraction, Activation Assay, Microarray, Gene Expression, shRNA, Sequencing, Control, Negative Control, Plasmid Preparation, Software

Journal: iScience

Article Title: Sulfonylurea receptor 1 deficiency alleviates acute lung injury by activating PARP1 in epithelial cells

doi: 10.1016/j.isci.2026.117627

Figure Lengend Snippet:

Article Snippet: Histone H3 , Proteintech Group , Cat# 17168-1-AP; RRID: AB_2716755.

Techniques: Cell-Signaling, Recombinant, Transfection, Caspase-3 Activity Assay, Protein Extraction, Knock-Out, Sequencing, Control, Plasmid Preparation, Software, Analysis

Journal: iScience

Article Title: IPO5 contributes to cetuximab resistance in colorectal cancer by modulating Ras signaling

doi: 10.1016/j.isci.2026.117714

Figure Lengend Snippet: The overexpression of IPO5 attenuates the cetuximab-mediated inhibition of Ras signaling (A) Subcellular fractionation and densitometric quantification of RASAL2 in cytoplasmic (Cyt) and nuclear (Nuc) fractions of RKO and LoVo cells following IPO5 knockdown or overexpression (lamin B and GAPDH served as fraction controls). (B) GST-Raf1-RBD pull-down assay assessing active Ras-GTP levels in CRC cells upon IPO5 modulation. (C) Rescue GST pull-down assays evaluating Ras-GTP levels in Caco2 and LoVo cells co-transfected with IPO5 and wild-type (WT) or NLS-mutant (Mut) RASAL2. (D) Western blot analysis and quantification of downstream proteins (p-AKT, AKT, p-MEK, MEK1/2, p-ERK, and ERK1/2) in cetuximab-treated cells under IPO5 modulation. (E) CCK-8 cell viability curves of Caco2 and LoVo cells treated with cetuximab following co-transfection of IPO5 with RASAL2-NLS-WT or NLS-Mut. Data are presented as mean ± SD ( n = 3 independent experiments). Statistical significance was determined using an unpaired two-tailed Student’s t test (A and B), one-way ANOVA with Tukey’s test (C), or two-way ANOVA with Tukey’s test (D and E). Exact p values are indicated.

Article Snippet: An anti-GAPDH antibody (Proteintech, USA, #10494-1-AP) was used as a cytoplasmic loading control, and an anti-Lamin B antibody (Proteintech, USA, #23498-1-AP) served as a nuclear loading control.

Techniques: Over Expression, Inhibition, Cell Fractionation, Knockdown, Pull Down Assay, GST Pulldown Assay, Transfection, Mutagenesis, Western Blot, Analysis, CCK-8 Assay, Cotransfection, Two Tailed Test