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: Peroxisome-dependent and -independent functions of PEX19 in mouse embryonic stem cells

doi: 10.1016/j.isci.2026.117658

Figure Lengend Snippet:

Article Snippet: Rabbit anti-ESRRB polyclonal antibody , Proteintech , 22644-1-AP; RRID: AB_2879140.

Techniques: Cell-Signaling, Magnetic Beads, Virus, Recombinant, ROS Assay, Reverse Transcription, SYBR Green Assay, Staining, Plasmid Preparation, Flow Cytometry, Western Blot, Expressing, Control, Software

Journal: iScience

Article Title: Peroxisome-dependent and -independent functions of PEX19 in mouse embryonic stem cells

doi: 10.1016/j.isci.2026.117658

Figure Lengend Snippet: Deletion of Pex19 impairs the growth of mESCs (A) Pex19 deficiency reduces the growth of mESCs. Equal numbers of WT and Pex19 KO 46C mESCs were seeded and cultured in ESGRO medium, and cell counts were analyzed on day 3 ( n = 3 biological replicates/group; data are presented as mean ± SEM; Student’s t test, ∗∗ p < 0.01). (B) Pex19 deficiency reduces the single-cell self-renewal capacity of mESCs. WT and Pex19 KO mESCs were cultured for colony formation for 5 days ( n = 3 biological replicates/group; data are presented as mean ± SEM, Student’s t test, ∗ p < 0.05). (C) Pex19 deficiency increases cell death and apoptosis of mESCs ( n = 3 biological replicates/group, Student’s t test, ∗∗ p < 0.01, ∗∗∗ p < 0.001). (D) Adding back PEX19 restores the peroxisomal targeting of PEX14. Scale bars, 5 μm. (E–G) Adding back PEX19 rescues defective cell growth and elevated cell death in Pex19 KO mESCs. E and F, n = 3–4 biological replicates/group; data are presented as mean ± SEM, one-way ANOVA, ∗q < 0.05, ∗∗q < 0.01, ∗∗∗q < 0.001, n.s., not significant). (H) Top enriched GSEA pathways in Pex19 KO vs. WT mESCs. Transcriptomes of WT and Pex19 KO 46C mESCs from the RNA-seq dataset were analyzed using GSEA as described in . NES, Normalized Enrichment Score; FDR, False Discovery Rate. See also .

Article Snippet: Rabbit anti-PEX14 polyclonal antibody , Proteintech , 10594-1-AP; RRID: AB_2252194.

Techniques: Cell Culture, Single Cell, Colony Assay, RNA Sequencing

Journal: iScience

Article Title: Peroxisome-dependent and -independent functions of PEX19 in mouse embryonic stem cells

doi: 10.1016/j.isci.2026.117658

Figure Lengend Snippet:

Article Snippet: Rabbit anti-PEX14 polyclonal antibody , Proteintech , 10594-1-AP; RRID: AB_2252194.

Techniques: Cell-Signaling, Magnetic Beads, Virus, Recombinant, ROS Assay, Reverse Transcription, SYBR Green Assay, Staining, Plasmid Preparation, Flow Cytometry, Western Blot, Expressing, Control, Software

Journal: iScience

Article Title: Peroxisome-dependent and -independent functions of PEX19 in mouse embryonic stem cells

doi: 10.1016/j.isci.2026.117658

Figure Lengend Snippet: Deletion of Pex19 induces mitotic block in mESCs (A) Deletion of Pex19 impairs cell cycle progression and induces polynuclear mESCs. WT, Pex19 KO, and Pex19 KO + PEX19 46C mESCs were stained with PI for DNA content, then subjected to FACS analysis ( n = 3 biological replicates/group; data are presented as mean ± SEM). (B) Deletion of Pex5 in mESCs fails to induce detectable defects in the cell cycle. WT, Pex5 KO, and Pex19 KO 46C mESCs were stained with DAPI for DNA content, then subjected to FACS analysis. (C) Deletion of Pex19 but not Pex5 induces a new fraction of polynuclear mitotic cells. WT, Pex5 KO, and Pex19 KO 46C mESCs treated with or without 50 ng/mL nocodazole were stained with an anti-H3pS10 antibody for detection of mitotic cells and DAPI for DNA content, then subjected to FACS analysis. The M-B population represents polynuclear mitotic cells arrested by deletion of Pex19 or treatment with nocodazole. (D and E) Deletion of Pex19 delays cell cycle progression of mESCs. WT and Pex19 KO 46C mESCs treated with 50 ng/mL nocodazole for 5 h were transferred into normal culture medium without nocodazole, and cell cycle progression was analyzed 15 and 25 min later (three groups of cells (80–250 cells/group) from randomly selected fields were analyzed for each time point for both WT and Pex19 KO mESCs; data are presented as mean ± SEM). Scale bars, 10 μm. See also .

Article Snippet: Rabbit anti-PEX5 polyclonal antibody , Novus , NBP1-87185; RRID: AB_11023925.

Techniques: Blocking Assay, Staining, Analysis

Journal: iScience

Article Title: Peroxisome-dependent and -independent functions of PEX19 in mouse embryonic stem cells

doi: 10.1016/j.isci.2026.117658

Figure Lengend Snippet:

Article Snippet: Rabbit anti-PEX5 polyclonal antibody , Novus , NBP1-87185; RRID: AB_11023925.

Techniques: Cell-Signaling, Magnetic Beads, Virus, Recombinant, ROS Assay, Reverse Transcription, SYBR Green Assay, Staining, Plasmid Preparation, Flow Cytometry, Western Blot, Expressing, Control, 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

Figure Lengend Snippet:

Article Snippet: Anti-p65 rabbit antibody , Proteintech , 10745-1-AP; RRID: AB_2178878.

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

Journal: Advanced Science

Article Title: Enzyme‐Linked Cycloaddition Assay (ELCA) for Rapid, Ultra‐Sensitive Monitoring of Secreted Sialoglycoproteins

doi: 10.1002/advs.78315

Figure Lengend Snippet: Quantifying secreted glycoproteins under standard culture conditions. (A) Schematic of the ELCA workflow. Supernatant samples containing azide‐modified glycoproteins are collected from metabolically labeled cells, adsorbed onto microwell plates, and subsequently labeled via strain‐promoted cycloaddition. Detection and quantification are performed using HRP. (B) XTT assay quantifying cellular metabolic activity under culture conditions with varying serum concentrations. ( n = 8). (C) ELCA analysis of supernatants from cells cultured under varying serum concentrations using standard colorimetric detection ( n = 4). (D, E) Signal‐to‐noise ratio (S/N) of ELCA signal comparing standard culture conditions (DMEM + 10% serum; (D) and serum‐free Opti‐MEM (E) at 24 h and 72 h time points, measured by standard colorimetric detection ( n = 4). (F) S/N of ELCA signal from cells cultured under standard conditions at 24 h and 72 h time points, measured using high‐sensitivity luminescent detection ( n = 4). (G, H) ELCA signal following treatment with 3 µ m Brefeldin A (BFA) for 24 h under serum‐free (G) or standard culture conditions (H), measured using high‐sensitivity luminescent detection ( n = 4). – (gray bars): blank control. (I–K) ELCA measurements of total glycoprotein secretion (I), Sandwich‐ELCA measurements of glycosylated GM‐CSF secretion (J), and ELISA measurements of total GM‐CSF secretion (K) following PMA stimulation in cells cultured under standard conditions for 6–48 h, using high‐sensitivity luminescent detection (I; n = 3) or standard colorimetric detection (J–K; n = 3). All assays were conducted in A375 cells. Statistical significance was assessed using a two‐tailed Welch's t ‐test (two conditions), one‐way ANOVA with Dunnett's multiple comparisons test (>2 conditions, 1 variable), or two‐way ANOVA with Tukey's multiple comparisons test (<2 conditions, 2 variables). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For detection, the plate was incubated with 0.5 μg/mL GM‐CSF polyclonal antibody (17762‐1‐AP, Proteintech) for 1 h at room temperature followed by 40 ng/mL HRP‐conjugated Goat Anti‐Rabbit IgG (SA00001‐2, Proteintech) for 1 h at room temperature.

Techniques: Modification, Metabolic Labelling, Labeling, XTT Assay, Activity Assay, Analysis, Cell Culture, Sensitivity, Control, Enzyme-linked Immunosorbent Assay, Two Tailed Test