Journal: iScience

Article Title: CGF induces ROS-mediated metabolic reprogramming and mitochondrial dysfunction to suppress colorectal cancer progression

doi: 10.1016/j.isci.2026.115273

Figure Lengend Snippet: CGF’s effect on cell cycle and apoptosis in CRC (A) Flow cytometry was used to analyze how CGF affects the cell cycle of HCT116 and HT29 cells at certain concentrations, with the percentage of cells in G1, S, and G2 phases shown in each panel. (B) Western blot analysis of the changes in cell cycle-related proteins CDK1, p-CDK1, and cyclin B1 in HCT116 and HT29 cells after CGF treatment. (C) RT-qPCR analysis of the relative expression levels of PUMA and NOXA genes in HCT116 and HT29 cells treated with different concentrations of CGF. (D) Western blot analysis of the changes in apoptosis-related proteins BCL2, PUMA, Noxa, C-caspase 9, and C-caspase 3 in HCT116 and HT29 cells after CGF treatment. (E) Flow cytometry was used to analyze apoptosis in HCT116 and HT29 cells treated with CGF. On the left is a representative plot showing apoptosis, utilizing Annexin V-FITC and PI double staining. Right: Analysis of early and late apoptosis in cells from each group using quantitative methods. (A–C and E) Data presentation is in the form of mean ± SEM. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb , CST , Cat # 9664; RRID:AB_2070042.

Techniques: Flow Cytometry, Western Blot, Quantitative RT-PCR, Expressing, Double Staining

Journal: iScience

Article Title: CGF induces ROS-mediated metabolic reprogramming and mitochondrial dysfunction to suppress colorectal cancer progression

doi: 10.1016/j.isci.2026.115273

Figure Lengend Snippet: The effect of CGF-induced ROS on the MAPK/ERK 1/2/c-MYC signaling pathway (A) Western blotting was used to analyze the expression of MEK, P-MEK, ERK 1/2, P-ERK 1/2, and c-MYC proteins in HCT116 and HT29 cells after treatment with specified CGF concentrations. (B) Immunofluorescence staining of HCT116 and HT29 cells after CGF treatment, showing changes in p-ERK (green). PMA was used to promote nuclear entry of p-ERK (200 nM, 4 h), and DAPI was used to stain the nucleus (blue), with merged images shown (scale bars, 10 μM). (C) Western blot was used to analyze the levels of MEK, P-MEK, ERK 1/2, P-ERK 1/2, and c-MYC proteins in HCT116 and HT29 cells following treatment with CGF (40 μM, 24 h) and NAC (500 μM, 24 h). (D) Untargeted metabolomics analysis of glycolytic metabolites after CGF treatment in HCT116 cells. Metabolites marked in blue represent those inhibited by CGF. (E and F) ELISA analysis of lactate secretion in the cell supernatant of HCT116 (E) and HT29 (F) cells after CGF treatment. (G and H) Analysis of mRNA expression levels of genes related to glycolysis in HCT116 and HT29 cells exposed to certain concentrations of CGF using RT-qPCR. (I) Western blot was used to analyze the expression levels of apoptosis-associated proteins (BCL2, PUMA, NOXA, C-caspase 9, and C-caspase 3) and EMT markers (N-cadherin and vimentin) in HCT116 and HT29 cells after they were treated with CGF. (J) Western blot analysis of the reversal of EMT markers N-cadherin, vimentin, SLUG, and SNAIL proteins in HCT116 and HT29 cells overexpressing c-MYC. (E–J) Data presentation is in the form of mean ± SEM. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Cleaved Caspase-3 (Asp175) (5A1E) Rabbit mAb , CST , Cat # 9664; RRID:AB_2070042.

Techniques: Western Blot, Expressing, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR