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. 2019 Apr 1;9(4):730–739.

Figure 4.

Figure 4

HMGB1 regulates TfR1 expression in the RAS-JNK/p38-dependent pathway. A. HL-60/NRASQ61L cells were lysed after treatment with erastin (5 μM) and/or Fer-1 (1 μM), then TfR1 expression was verified by western blot. B. HL-60/NRASQ61L cells were transfected with TfR1 siRNA and control siRNA, and TfR1 expression was verified by western blot. Then the two groups of cells were stimulated with erastin at the indicated doses for 48 h. Cell viability was assayed. Ctrl, control. (n = 3, *P < 0.05 versus the control group). C. HL-60/NRASQ61L cells were treated with erastin (5 μM) combined with SP600125 (10 μM) and SB202190 (10 μM) for 48 h. TfR1 expression and the phosphorylation of p38 (p-P38) and JNK1/2 (p-JNK1/2) were assayed by western blot. D. HL-60/NRASQ61L cells were transfected with NRAS siRNA and control siRNA vector, and NRAS expression was verified by western blot. Then two groups of cells were stimulated with erastin (5 μM) for 48 h. TfR1, p-P38 and p-JNK1/2 were assayed by Western blot. Ctrl, control. E. HL-60/NRASQ61L cells were transfected with HMGB1 shRNA (HMGB1 shRNA1 and HMGB1 shRNA2) and control shRNA vector and then stimulated with erastin (5 μM) for 48 h. TfR1, p-P38 and p-JNK1/2 expression levels were assayed by western blot. F. HL-60/NRASQ61L cells were transfected with HMGB1 plasmid and empty vector, and HMGB1 expression was verified by western blot. Then, two groups of cells were stimulated with erastin (5 μM) combined with SP600125 (10 μM) and SB202190 (10 μM) for 48 h. TfR1 expression was assayed by western blot. Necrostatin-1 pretreatment was used in all experiments. All experiments were conducted in triplicate, and the data are presented as the mean ± SD. Fer-1, ferrostatin-1.