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. 2019 Mar 1;9(3):546–561.

Figure 2.

Figure 2

SPHK2 regulates cholangiocarcinoma cell apoptosis via NOXA. A. Cells were transfected with 50 nM Control siRNA (siCon) or NOXA siRNA (siNOXA) for 48 h and then treated with 50 μM ABC294640 for 48 h. NOXA mRNA level was analyzed by Real-time qPCR. B. Cells were transfected with 50 nM Control siRNA (siCon) or NOXA siRNA (siNOXA) for 48 h and then treated with 50 μM ABC294640 for 24 h. NOXA protein level was analyzed by Western immunoblotting. C-E. Cells were transfected with 50 nM Control siRNA (siCon) or NOXA siRNA (siNOXA) for 48 h and then treated with 50 μM ABC294640 for 72 h. Cell viability was determined by CCK-8 assay. Apoptosis was analyzed by PARP cleavage through Western immunoblotting in both cell lines and by Annexin V-FITC/PI labeling followed by flow cytometry in RBE cells. F. Kaplan-Meier curves showing the overall survival rate of 36 patients in the TCGA dataset according to the expression status of NOXA. G. Correlation between SPHK2 mRNA expression and NOXA mRNA expression in a cholangiocarcinoma dataset (GSE26566). The r and P values were determined by Pearson correlation analysis. Quantitative analysis from 3 independent experiments (one-way ANOVA with a Turkey post hoc test; data are shown as mean ± SEM; *P < 0.05, **P < 0.01) are shown.