Fig. 5

CCT3 upregulation of slc7a11 inhibits ferroptosis. A qRT-PCR analysis of GPX4 and slc7a11 in Ctrl and CCT3 overexpressed H1975 cells, and in shCtrl, shCCT3#1 and shCCT3#2 H2228 cells. GAPDH is the internal control. *p < 0.05. B Western blot analysis of slc7a11 in cells as shown in A. GAPDH is the internal control. *p < 0.05. **p < 0.01. The blots were cut prior to hybridisation with antibodies during blotting. C Dual luciferase activity was checked in H2228 cells transfected with shRNAs (shCtrl, shCCT3#1 and shCCT3#2) or in H1975 cells transfected with pCDNA3.1 plasmids (pCDNA3.1-Ctrl or pCDNA3.1-CCT3), pGL3.Basic-slc7a11 promoter and TK. D Sperman correlation between CCT3 and slc7a11 in LUAD patients was analyzed from TCGA. Overall survival of LUAD patients (slc7a11 high and low expression groups) was analyzed from TCGA. E qRT-PCR analysis of slc7a11 and CCK8 analysis of cell proliferation were performed in Ctrl, CCT3 and CCT3 + sislc7a11 H1975 cells. *p < 0.05. **p < 0.01. F qRT-PCR analysis of slc7a11 and CCK8 analysis of cell proliferation were performed in shCtrl, shCCT3 and shCCT3 + slc7a11 H2228 cells. *p < 0.05. **p < 0.01. G and H Apoptosis and ferroptosis were detected by using Annexin V-FITC/PI staining in Ctrl, CCT3 and CCT3 + sislc7a11 H1975 cells (G), and in shCtrl, shCCT3 and shCCT3 + slc7a11 H2228 cells. Left, representative images of apoptosis and necroptosis. Right, quantification results. *p < 0.05. **p < 0.01