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. 2023 Sep 4;9:331. doi: 10.1038/s41420-023-01630-8

Fig. 2. In vitro assays confirmed that CRTC3 significantly induced resistance to IFN-γ treatment in HCC.

Fig. 2

a Expression levels of CRTC3 mRNA in tumor and normal tissues were revealed by analysis of the TCGA-LIHC database. b Kaplan–Meier curves did not reveal significant correlations between CRTC3 expressions and HCC patient survival outcomes. c Western blotting confirmed the knockout efficacies of CRTC3 in HepG2 and Hep3B cell lines. d Flow cytometry analyses were performed to reveal cell death under conditions of CRTC3 knockout and IFN-γ treatment (100 ng/mL). CRTC3 knockout alone did not have significant effects on cell death. Simultaneous treatment with IFN-γ induced CRTC3-KO HepG2 and Hep3B cell deaths, compared to the control (n = 3). e Colony formation assays were used to assess long-term cell viability after treatment with IFN-γ (100 ng/mL; n = 3). f Caspase 3/7 assay demonstrated comparable caspase activities induced by IFN-γ (100 ng/mL) in CRTC3-WT and CRTC3-KO HCC cell lines (n = 3). However, this effect was not enhanced by CRTC3 knockout. *P < 0.05; **P < 0.01; ***P < 0.001; Student’s t test.