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. 2023 May 3;42(24):2007–2016. doi: 10.1038/s41388-023-02706-6

Fig. 4. Everolimus activates the extrinsic pathway in human colonic epithelial cells upon APC loss.

Fig. 4

Human NCM356 cells were transfected with control or the indicated siRNA duplexes and treated with Everolimus (10 μm) for the indicated times. A Attached cells at 48 h were visualized by crystal violet staining. B Apoptosis was measured at 48 h by Hoechst staining of fragmented nuclei. C qRT-PCR analysis of the indicated genes at 24 h. D Western blots of the indicated proteins at 2 h. E Western blots of the indicated proteins at 24 h and F Apoptosis at 48 h measured by Hoechst staining of fragmented nuclei with Myc siRNA. G qRT-PCR analysis of the indicated genes at 24 h with Myc siRNA. H Western blots of the indicated proteins at 24 h and I apoptosis at 48 h measured by Hoechst staining of fragmented nuclei with BID siRNA. Arrows indicated cleaved bands. B, C, F, G, I, n = 3 independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 (Student’s T-Test, two-tailed).