Fig. 5

HIGD1A transcriptionally activates c-Myc expression to regulate the cancer phenotype of HCC cells. A ChIP assay confirming that c-Myc is transcriptionally activated by HIGD1A. Representative electrophoresis images (left) and RT-PCR (right) results for the ChIP assay. B Luciferase reporter assay was performed in HepG2 and MHCC97H cells infected with shCtrl or shHIGD1A lentivirus and transfected with pcDNA3.1 or pcDNA3.1-HIGD1A, respectively. C Cell viability of c-Myc-overexpressing HepG2 and MHCC97H cells infected with shCtrl or shHIGD1A lentivirus as measured by CCK8 assay at the indicated time points. D Western blotting analysis indicating the expression of c-Myc and EMT markers, E-cadherin and N-cadherin in c-Myc-overexpressing HepG2 and MHCC97H cells infected with shCtrl or shHIGD1A lentivirus. E and F Transwell assay showing that c-Myc overexpression reversed shHIGD1A-induced inhibition of cell migration in HepG2 and MHCC97H cells. G and H Transwell assay with Matrigel demonstrated c-Myc overexpression reversed shHIGD1A-induced inhibition of cell invasion in HepG2 and MHCC97H cells. Results shown are mean ± SEM. *P <0.05, **P <0.01, ***P <0.001. Scale bar, 100µm