Figure 3.

The effects of KARS knockdown on HCC cell lines. (A) Relative expression of KARS1 mRNA in HCC cell lines compared to SNU398 (n = 4‐6). (B) KARS protein expression in HCC cell lines. (C) Protein expression levels of KARS and cleaved‐Caspase 3 in Huh7 and SNU398 HCC cells following shRNA mediated knockdown of KARS. The intensity was quantified relative to β‐actin (n = 4‐8). (D) KARS1 mRNA expression following shRNA mediated knockdown quantified by qRT‐PCR (n = 4‐10). (E) Effects of KARS knockdown on cell growth measured by MTT assay following 1, 3 and 6 days of culture (n = 11, 22). (F) KARS knockdown affects the number of single cell formed colonies as assayed 2 weeks following seeding (n = 5). (G) The effect of KARS knockdown on cell cycling was measured by Propidium Iodide staining, and the fraction of cells in G0/G1 was quantified (n = 4‐6). Quantification of C, D and F data of knockdown groups were relative to the shCTR group. Data are presented as mean ± SEM. *P < .05; **P < .01; ***P < .001, by the Mann‐Whitney test. HLO, healthy liver organoids; HCC, hepatocellular carcinoma; FC, fold change; KARS1, Lysyl‐tRNA Synthetase