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

Additional file 1: Figure S1. Establishment and characterization of the acquired 5-FU-R CRC cell line models, related to Fig. 1. a. Acquired 5-FU-R CRC cell line models (HCT8, HCT15, and LoVo) were established by inducing WT CRC cells with a gradient increased 5-FU concentrations over a period of approximately 8 months. b. Phalloidin (green) and DAPI (blue) staining to visualize the cell morphology and nuclei. Scale bar = 50 μm. c. EdU incorporation assays of cell proliferation with or without 10− 5 M 5-FU treatment. EdU positive cells (red), nucleus (blue). Scale bar = 100 μm. d. Cell cycle distribution with or without 10− 5 M 5-FU treatment. e. Cell apoptosis with or without 10− 5 M 5-FU treatment. The four populations were distinguished as follows: viable cells (PE Annexin-V and 7-AAD negative), early apoptotic cells (PE Annexin-V positive and 7-AAD negative), late apoptotic cells (PE Annexin-V and 7-AAD positive), and dead cells (PE Annexin-V negative and 7-AAD positive). f. Comparing basal respiration, maximal respiration, and spare capacity in 5-FU-R CRC cells to WT CRC cells for assessing the mitochondrial respiration function. For all studies n was ≥3. Data are means ± SEM. Bar chart data were compared by Student’s t-test (* p < 0.05, ** p < 0.01, and *** p < 0.001).