Fig. 1

P300 was downregulated during ferroptosis of HASMCs. (A) Relative cell viability of HASMCs measured with a CCK8 kit after cystine deprivation (CD) treatment for 0, 4, 8, 12, 16, and 24 h (n = 6 per group). (B) Relative cell viability of HASMCs after treatment with different concentrations of imidazole ketone-erastin (IKE) (0, 0.5, 1, 2.5, 5, 10 µM) (n = 6 per group). C-F. The level of lipid ROS (oxidized BODIPY-C11/non-oxidized BODIPY-C11 ratio) detected by using a BODIPY-C11 kit in HASMCs after treatment with CD (C-D) and IKE (E-F) (n = 3 per group). G. Representative images of immunofluorescence staining with 4-HNE in HASMCs treated with DMSO and Fer-1 after CD and IKE stimulation for 16 h. H. Quantitative analysis of immunofluorescence staining of 4-HNE (n = 4 per group). I. The level of MDA measured by using an MDA assay kit in HASMCs treated with DMSO and Fer-1 after CD and IKE stimulation for 16 h. (n = 5 per group). J-M. P300 protein levels in HAMSCs after CD (J-K), IKE (L-M) stimulation for 16 h and treatment with Fer-1, 3-MA and Emricasan were measured by western blot (n = 4 per group). J and L. Representative western blot; K and M. Quantitative results. β-actin served as a loading control. Values are means ± SD; ***p < 0.001, **p < 0.01, NS, no significant