Fig. 5

Inhibiting GRIM-19 activated the STAT3/HIF-1α pathway and strengthened HCC progression. GRIM-19 knockdown plasmids and/or SATB2-AS1 overexpression plasmids were transfected into Huh7 cells. A, B: Colony formation experiment and EdU staining were utilized to detect cell proliferation. Scale bar = 50 μm. C: Western blot was conducted for detecting cell cycle-related proteins (including p21, Cyclin D1, CDK4, Cyclin E1, and CDK2) levels in HCC cells. D. Flow cytometry (FCM) was adopted to monitor the HCC cell apoptosis rate. E: Western blot was conducted for detecting apoptosis-related proteins (including Bad, bcl2, Bax, cleaved Caspase3) levels in HCC cells. F: Huh7 cell migration and invasion were tested by Transwell assay. Scale bar = 100 μm. G: WB was implemented to determine the levels of EMT-related proteins (E-cadherin, Vimentin, and N-cadherin). H: The GRIM-19/STAT3/HIF-1α expression in Huh7 cells was measured by WB. * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001. N = 3