cck 8 solution  (Dojindo Labs)

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: After 48 h of drug exposure, the medium was replaced with 110 μL/well of CCK-8 reaction solution (10 μL of CCK-8 reagent [Dojindo, Kumamoto, Japan] + 100 μL of RPMI-1640 medium).

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: 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: After 48 h of drug exposure, the medium was replaced with 110 μL/well of CCK-8 reaction solution (10 μL of CCK-8 reagent [Dojindo, Kumamoto, Japan] + 100 μL of RPMI-1640 medium).

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: After 48 h of drug exposure, the medium was replaced with 110 μL/well of CCK-8 reaction solution (10 μL of CCK-8 reagent [Dojindo, Kumamoto, Japan] + 100 μL of RPMI-1640 medium).

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