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. 2020 Oct 10;11(10):843. doi: 10.1038/s41419-020-03075-8

Fig. 7. USP7 and ZNF638 affect lipogenesis-associated hepatic carcinogenesis.

Fig. 7

A SK-Hep1 cells with or without stable ZNF638 knockdown, stable USP7 knockdown, P22077 treatment (5 μM), and C75 treatment (1 μg/ml) were cultured with or without fructose (8 mM) for 14 days; after that, the cell clones were harvested and calculated by ImageJ. B Seventy percent of different SK-Hep1 cells (ZNF638 knockdown, USP7 knockdown, treatment of 5 μM P22077, treatment of 1 μg/ml C75) and 30% of LXR cells were mixed and glued with Collagen I (5 μg/ml); the culture medium with or without fructose (8 mM) was added with 20 ng/ml b-FGF, 20 ng/ml HGF to promote the growth of both cell lines. At the time points of 3 days and 5 days, organoids volumes of different groups were analyzed. C SK-Hep1 cells with or without stable ZNF638 knockdown, stable USP7 knockdown, P22077 treatment (10 μM), and C75 treatment (2.5 μg/ml) were pre-scratched and cultured with or without fructose (8 mM) for 24 h, the migration area (area% of per field of view) of different groups was evaluated by ImageJ. D SK-Hep1 cells with or without stable ZNF638 knockdown, stable USP7 knockdown, P22077 treatment (10 μM), and C75 treatment (2.5 μg/ml) were cultured with or without fructose (8 mM) for 48 h in transwell chambers; the invaded cells were calculated and analyzed by ImageJ. E The overall survival difference between high and low expression of USP7 or ZNF638 in HCC patients was analyzed using the transcriptome data from TCGA; the cutoff value was computed by survminer R package. F The discrepancy in the ratio of HCC patients with different USP7/ZNF638 expression from different etiologies was analyzed based on the data from TCGA. G USP7 and ZNF638 expression were evaluated in four paired HCC paraffin sections (2 of steatosis, 2 of non-steatosis) by immunochemistry. The representative statistical results were performed using one- or two-way ANOVA test and shown as means ± SEM from three independent experiments. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.