Fig. 4

SCD is the downstream target of CREB1. A Selection of downstream genes of CREB1 by the intersection of ferroptosis-related genes, target genes of CREB1 screened by ChIP high-throughput data, and downregulated genes in CREB1-knockdown A549 cells revealed by RNA-seq. B Fifteen potential target genes of CREB1 with the most pronounced fold change in CREB1-knockdown A549 cells. C RT-qPCR revealed the mRNA expression of GCH1, CBS, SCD, and HIF1A between NC and CREB1-knockdown H1299 cells. D Pearson correlation between the SCD and CREB1 expression in NSCLC tissue from TCGA database. E, F RT-qPCR (E) and immunoblotting (F, cropped blots) revealed the SCD expression in NSCLC cells after transfected with vector or SCD-overexpression lentivirus. G RT-qPCR revealed the mRNA expression of SCD in A549 and H1299 cells after treatment with H89 (30 μM) or cAMP (100 μM) for 48 h. H SCD level in A549 and H1299 cells (cropped blots) after transfection with NC/ CREB1-SH1/ CREB1-SH2 lentivirus or treatment with H89 (30 μM)/ cAMP (100 μM) for 48 h. I, J, K RT-qPCR (I) and Immunoblotting (cropped blots, J & K) revealed the SCD expression with indicating treatments in A549 and H1299 cells. NSCLC, non-small cell lung cancer. Data were analyzed by two-way ANOVA or Student’s t-test and were presented by mean ± SD from three biological replicates. ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001