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. 2017 Jan;360(1):14–22. doi: 10.1124/jpet.116.236620

Fig. 3.

Fig. 3.

The role of NOX5-S in acid-induced SODD expression. (A) A typical image of three Western blot analyses and (B) summarized data showed that acid treatment significantly increased SODD protein expression in FLO cells, an increase that was significantly decreased by knockdown of NOX5 with its siRNA (N = 3). (C) Acid treatment significantly increased SODD mRNA expression in a Barrett’s cell line CP-A cells, an increase that was significantly decreased by knockdown of NOX5 with its siRNA (N = 3). (D) Acid treatment remarkably increased the luciferase activity of SODD promoter in FLO cells transfected with SODD reporter plasmid pGL3-SODDP (N = 3), which was generated by ligating a SODD promoter fragment (−1008 to 20 from ATG) into the pGL3-basic vector, indicating that acid treatment may activate SODD promoter. Acid-induced increase in the luciferase activity was significantly decreased by knockdown of NOX5 (N = 3), suggesting that acid-induced activation of SODD promoter may depend on the activation of NOX5-S. (E) Acid treatment significantly decreased cell apoptosis in FLO EA cells (N = 3–5). (F) Acid-induced decrease in caspase 9 activity was significantly inhibited by knockdown of SODD, suggesting that SODD may mediate acid-induced decrease in cell apoptosis in FLO cells. (G) Acid-induced decrease in cell apoptosis was significantly inhibited by knockdown of SODD, suggesting that SODD may mediate acid-induced decrease in cell apoptosis in FLO cells. Percentage of decrease was calculated as follows: % decrease = (control-acid) × 100/control. Analysis of variance, *P < 0.02, ##P < 0.01, and ***P < 0.0001, compared with control siRNA group; **P < 0.01, compared with control siRNA + acid group; ▴▴P < 0.001, compared with control siRNA + acid group. t test, #P < 0.05; ▴P < 0.0001.