Skip to main content
Fig. 1 | Journal of Experimental & Clinical Cancer Research

Fig. 1

From: ROS/PI3K/Akt and Wnt/β-catenin signalings activate HIF-1α-induced metabolic reprogramming to impart 5-fluorouracil resistance in colorectal cancer

Fig. 1

Establishment and characterization of the acquired 5-FU-R CRC cell line models. a CCK8 assays to assess 5-FU sensitivity of 5-FU-R CRC cells compared with WT CRC cells, and cells cultured with increasing concentrations of 5-FU for 72 h. All experiments were performed with 6 replicates. b Proportion of EdU-positive cells with or without 10− 5 M 5-FU treatment. c Cell cycle data for G1 phase, G2 phase and M phase distribution with or without 10− 5 M 5-FU treatment. d Western blots of cell cycle markers. β-Actin was used as the internal reference. e Apoptosis with or without 10− 5 M 5-FU treatment. Both early and late apoptotic cells are shown. f Western blots of apoptosis markers. β-Actin was used as the internal reference. g Mitochondria marked by Mito-tracker (Texas Red). Data are mean MFI (Texas Red channel) ± SEM. h Typical images of the ultrastructure of mitochondria by transmission electron microscopy (TEM). Scale bar = 5 μm/1 μm. i Western blots of five enzymes of the mitochondrial respiratory chain. β-Actin was used as the internal reference. j RT-qPCR analysis of mitochondrial enzyme genes in 5-FU-R CRC cells relative to WT CRC cells. ACTB was used as the internal reference. k 5-FU-R and WT cells were distributed into in 96-well Seahorse assay plates and oligomycin (1.5 μM), FCCP (1 μM) and rotenone/antimycin A (0.5 μM) were successively added to measure OCR. l ATP production of 5-FU-R CRC cells relative to WT CRC cells detected by fluorometric analysis. ATP production was normalized to total protein concentration. For all studies n was ≥3. Data are means ± SEM. Bar chart data were compared by Student’s t-test or ANOVA (ns = not significant, * p < 0.05, ** p < 0.01, and *** p < 0.001)

Back to article page