Extended Data Fig. 6: CKB suppresses ferroptosis in a GPX4 pS104 dependent manner.
From: Creatine kinase B suppresses ferroptosis by phosphorylating GPX4 through a moonlighting function
(a-l) Data are the mean ± SD (n = 3), *P < 0.01; **P < 0.001; N.S., not significant by two-tailed Student’s t-test. (a-d) Huh7 and HCCLM3 cells expressing GPX4 shRNA with reconstituted expression of the indicated GPX4 proteins were treated with or without cystine deprivation (a, c) or 20 μM Erastin (b, d) for 24 h. Lipid ROS-positive cells (a, b) and cell death (c, d) were measured respectively. (e, f) Huh7 and HCCLM3 cells expressing GPX4 shRNA with reconstituted expression of the indicated GPX4 proteins were treated with different doses of sulfasalazine (SAS) (e) or Erastin (f) for 24 h, cell viability were measured. (g-l) Parental Huh7 cells and the indicated clones with knock-in expression of GPX4 S104A mutants were stably transfected with CKB shRNA and reconstituted with indicated CKB proteins. The cells were treated with or without cystine deprivation (g, i) or 20 μM Erastin (h, j) for 24 h. Lipid ROS-positive cells (g, h) and cell death (i, j) were measured respectively. The cells were treated with different doses of sulfasalazine (SAS) (k) or Erastin (l) for 24 h, cell viability were measured.
