Fig. 4

LPCAT1 is a direct transcriptional target of NRF1. A-B NRF1 function was inferred by GO (A) and KEGG (B) enrichment analysis with top 3000 putative NRF1 target genes. C Venn plot showed the intersection of the three gene sets (top 3000 NRF1 putative target genes, top 500 most differential survival genes in HCC and phospholipid biosynthetic process gene set). (D) Upper: screenshot from ENCODE website showing the NRF1 binding peak in ChIP-seq experiment near LPCAT1 transcription start site. Lower: the putative binding sites of NRF1 on LPCAT1 promoter predicted by JASPAR website was shown (P1:55 ~ 65;P2:9 ~ 19;P3:-16 ~ -6;P4:-157 ~ -147;P5:-324 ~ -314;P6:-695 ~ -685). E–F The mRNA and protein levels of LPCAT1 in Huh7 and MHCC97H after NRF1 knockdown or overexpression were determined with qRT-PCR (E) and Western blot (F), respectively. G ChIP-qPCR assay was performed to analyse the binding of NRF1 on LPCAT1 promoter. α-NRF1, anti-NRF1 antibody. H Dual luciferase reporter assay showed the relative LPCAT1 promoter luciferase activity in Huh7 cells after NRF1 overexpression. *p < 0.05, ** p  < 0.01, *** p  < 0.001