Fig. 4

HIGD1A knockdown causes cell cycle arrest in the G2/M phase through downregulating c-Myc target genes. A Transcriptome strategy of RNA-seq conducted on HepG2 cells infected with shCtrl or shHIGD1A lentivirus for 48 h. Each group contained three biological replicates. Volcano plots of differential gene expression in shCtrl and shHIGD1A infected HepG2 cells. B Signaling pathway enrichment in different groups analyzed by GSEA. Bubble plot shows the main decreased molecular hallmarks (GSEA) enriched in the shHIGD1A groups. Normalized enrichment scores (NES) across gene sets are shown. C mRNA levels of HIGD1A, c-Myc, ODC1, and markers of G2/M checkpoints in HepG2 cells infected with shCtrl or shHIGD1A lentivirus. β‐actin was used as a loading control. D Protein levels of HIGD1A, c-Myc, and ODC1 in HepG2, Huh7, and MHCC97H cells infected with shCtrl or shHIGD1A lentivirus. E Left: HepG2, Huh7, and MHCC97H cells treated with shCtrl or shHIGD1A were synchronized with APH for 24 h. After the removal of APH, HepG2, Huh7, and MHCC97H cells were stained with PI and analyzed by flow assay. Right: proportion of cell populations in G1/S and G2/M phases. Results shown are mean ± SEM. *P <0.05, **P <0.01, ***P <0.001