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. 2021 Feb 8;8(8):2002874. doi: 10.1002/advs.202002874

Figure 2.

Figure 2

OTUD1 activates the caspase‐independent apoptotic pathway. A,B) Representative images and fraction of EdU‐positive cells. Scale bars, 60 µm. The data are the means ± s.e.m.; n  =  3. Two‐tailed t tests, ***p < 0.001, **p < 0.01, ns: not significant. C) Representative image and tumor weights of xenografts from KYSE30 cells expressing EV, OTUD1, or OTUD1C320A and treated with saline or DDP. The data are the means ± s.e.m.; n = 7. Two‐tailed t tests, ***p < 0.001, **p < 0.01. D) Representative image and tumor weights of xenografts from KYSE150 cells expressing shC, shD1#1, or shD1#2 shRNAs and treated with saline or DDP. The data are the means ± s.e.m.; n = 7. Two‐tailed t tests, ***p < 0.001, ns: not significant. E,F) Statistical analyses of the cell death fraction of the indicated cells as determined using flow cytometric analysis. The data are the means ± s.e.m.; n  =  3. Two‐tailed t tests, ***p < 0.001, **p < 0.01, ns: not significant. G) Representative image and statistical analyses of JC‐1 staining of KYSE30 cells expressing EV, OTUD1 or OTUD1C320A and treated with DDP. Scale bars, 30 µm. Fluorescence intensity (fold) indicates the relative intensity of Red/Green. The data are the means ± s.e.m.; n  =  5. Two‐tailed t tests, **p < 0.01, ns: not significant. H) In vitro growth of KYSE30 cells expressing EV or OTUD1 and treated with or without Z‐VAD (50 × 10−6 m). The data are the means ± s.d.; n = 5. Two‐tailed t tests, ***p < 0.001. I) Cell death fractions of the indicated KYSE30 cells expressing EV or OTUD1 and treated with DDP together with or without Z‐VAD (50 × 10−6 m). The data are the means ± s.e.m.; n  =  3. Two‐tailed t tests, **p < 0.01, *p < 0.05. J) Representative image and statistical analyses of JC‐1 staining of KYSE30 cells expressing EV or OTUD1 and treated with DDP or Z‐VAD (50 × 10−6 m). Scale bars, 30 µm. Fluorescence intensity (fold) indicates the relative intensity of Red/Green. The data are the means ± s.e.m.; n  =  5. Two‐tailed t tests, **p < 0.01.