Fig. 4: LDHB regulates GSH-dependent ferroptosis defense through SLC7A11.

a Heat maps showing the mRNA fold change of ferroptosis-related genes (n = 25) in siLDHB cells (KD) compared to siNT cells (WT) (based on RNA-seq data). b, c Immunoblots of the indicated cells transfected for 48 h with siNT (-) or siLDHB (+) or stably expressing shRNAs. d Immunoblot of A549 stably expressing shNT or shLDHB were further transduced with either an empty vector or a SLC7A11-expressing plasmid (pCMV-SLC7A11). e, f Viability and lipid ROS assay of the indicated cells treated with erastin (A549, 5 µM; H838, 2.5 µM) or sulfasalazine (SSZ; A549, 0.25 mM; H838, 1 mM) for 24 h. g Immunoblots of A549 transfected (48 h) with siLDHB or siNT and further treated (16 h) with erastin (5 uM) or Sulfasalazine (1 mM). Murine KP cells expressing Ldhb shRNA or control shRNA were treated with erastin (10 uM) or Sulfasalazine (1 mM) for 24 h. h, i Immunoblots h and viability assay i of A549 cells transfected with siLDHB, siSTAT1 or siNTs for 24 h, followed by further treatment with erastin (10 µM) for another 24 h. j SLC7A11 mRNA fold change in A549 cells transfected (72 h) with siSTAT1 compared to A549 cells transfected with siNT. **p < 0.01 by Student’s t-test. k ChIP of STAT1 in A549 cells transfected with siLDHB or siNT, or treated with IFNγ. STAT1 binding to the SLC7A11 promoter (GAS2 domain) was quantified by qPCR. Results are expressed as fold change in site occupancy over IgG control and shown as mean ± SD from three independent experiments (n = 3). **p < 0.01 by two-way ANOVA.