Circ-DENND4C regulated HCC cell growth, invasion and stemness.
a qRT-PCR examined circ-DENND4C
expression in HCC cells and normal liver epithelial cells. b, c Gel
electrophoresis and ActD treatment verified circular characteristic of
circ-DENND4C. d qRT-PCR verified
knockdown efficiency of sh-circ-DENND4C in HCC cells. e, f.
Immunofluorescence staining assay (scale bar = 30 μm) and colony
formation assay detected the proliferation of HCC cells transfected with
sh-circ-DENND4C#1/2. g Cell cycle was
analyzed by flow cytometry upon circ-DENND4C knockdown. h TUNEL assay (scale bar = 100 μm) revealed HCC
cell apoptosis with transfection of sh-circ-DENND4C#1/2. i The protein levels of Cyclin D1, CDK4, Bcl-2
and Bax were determined through western blot analysis. j Transwell assay (scale bar = 100 μm) detected
cell invasion in HCC cells transfected with sh-circ-DENND4C#1/2.
qRT-PCR and western blot detected mRNA and protein levels of stemness
biomarkers in HCC cells upon circ-DENND4C depletion. *P < 0.05,
**P < 0.01