Fig. 1

m⁶A modification is increased during ferroptosis cell death in CRC. (A-B) CRC cells treated with 5 µM, 10 µM, and 20 µM of Erastin (or indicate concentration of RSL3), and then harvested cells for counting cell number at indicated day 1, day 2, day 3, and day 4 to determine the cell proliferation. (C-D) CRC cells pr-treated with Erastin (or RSL3) for 4 h, subsequently treated with or without of DFO (DFO; 100 nM), 3MA (3MA; 0.2 mM), Z-VAD-FMK (Z-VAD; 1 µM), or ferrostatin1 (Fer-1; 100 nM) for another 72 h, and then harvested cells for counting cell number to determine the cell proliferation. (E-F) CRC cells treated with 5 µM, 10 µM, and 20 µM of Erastin (or 200 nM, 400 nM, and 800 nM of RSL3) for 72 h, and then the total RNA were harvested for ELISA to determine the m⁶A levels. (G-H) CRC cells treated with 20 µM of Erastin (or 800 nM of RSL3) for 4 h, subsequently treated with or without DFO (100 nM) for another 72 h, and then the total RNA were harvested for ELISA to determine the m⁶A levels. (I-J) CRC cells treated with 20 µM of Erastin (or 800 nM of RSL3) (4 h) in the absence or presence of Fer-1 (100 nM) for 72 h, and then the total RNA were harvested for ELISA to determine the m⁶A levels. (All error bars, mean values ± SEM (standard error of mean), p values were determined by unpaired two-tailed Student’s t test of n = 3 independent biological experiments. *p < 0.05; **p < 0.01; ***p < 0.001)