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. 2020 Jul 22;9(8):1760. doi: 10.3390/cells9081760

Figure 3.

Figure 3

Monitoring apoptosis induced by marine compounds using flow cytometry in Jurkat and HCT116 cells. (A) Jurkat WT, Cas-8-/- and FADD-/- cells were treated with the indicated compounds for 24 h then stained with 7AAD and Annexin V and fluorescence was analyzed by flow cytometry. (B) Flow cytometry and Western blot analysis to determine the sensitivity of HCT116-Cas8-/- toward TRAIL-and FasL-induced cell death. Cells were treated with and without TRAIL and FasL for 6 h then subjected to dual staining as above and apoptotic percentage was determined by flow cytometry. For Western blot, cell lysates were prepared and loaded on SDS-PAGE (12%) followed by immunoblotting using the corresponding antibodies. HSC70 was used as loading control. (C) Colon HCT116 (WT, Cas8-/-, and TRAIL-R1/TRAIL-R2-/-) cancer cell lines were treated as previously mentioned. All values are presented here as ±SD (n = 3). Significance was evaluated by two-way ANOVA. *p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001.