Loss-of-function mutations of microRNA-142-3p promote ASH1L expression to induce immune evasion and hepatocellular carcinoma progression

Figure 4 The expression of ASH1L in MHCC-97H and knockdown cell lines and proliferation assay, migration and invasion assay of 97H-ASH1L-knockdown. A: Fluorescence imaging was conducted post-lentiviral transfection to visualize the ASH1L protein levels and western blot analysis was performed to compare the protein expression between MHCC-97H Scrambled (Scr) and Knockdown (KD) groups; B: The quantitative analysis of the western blot results provided a numerical comparison of ASH1L protein expression between the MHCC-97H Scr and KD groups; C: Quantitative real-time polymerase chain reaction (RT-qPCR) analysis was utilized to measure the differences in ASH1L gene expression between MHCC-97H Scr and KD groups, providing insights into the transcriptional effects of ASH1L knockdown; D: The colony formation assay indicated that the downregulation of endogenous ASH1L in MHCC-97H cells led to a decrease in the average number of colonies formed, suggesting a reduced clonogenic capacity; E: Cell counting kit 8 assays were employed to assess cell proliferation, and the results demonstrated that the downregulation of endogenous ASH1L significantly decreased the growth rate of MHCC-97H cells; F: Transwell chamber assays were conducted to evaluate and compare the abilities of MHCC-97H Scr and KD cells in terms of migration and invasion, revealing the impact of ASH1L on these cellular behaviors; G: Quantitative analyses of the Transwell assay data statistically compared the migration and invasion capabilities of MHCC-97H Scr and KD cells, further elucidating the role of ASH1L in cell movement and invasion; H: Cell apoptosis was detected by flow cytometry, and the results showed the influence of ASH1L knock-out on apoptosis. ^aP < 0.05. ^bP < 0.01. ^cP < 0.001. ^dP < 0.0001. GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; Scr: Scrambled; APC: Allophycocyanin conjugate; PE: Phycoerythrin; OD: Optical density.