Figure 7.

Effects of treatment with Doxo alone and in combination with radicicol on iNOS (A) gene and (B) protein expression, and (C) nitrotyrosine formation, and with the (D) addition of L-NAME for apoptosis, of the H9c2 cells. Doxo (1 µM) was administered for 3 or 6 h, and where indicated, radicicol (1 µM) was administered 30 min prior to Doxo. (A) iNOS mRNA levels analyzed by RT-qPCR, with the data normalized to the HPRT1 gene The data are presented as the means ± SD for at least 3 independent experiments, which each performed in triplicate. (B) iNOS expression was detected by western blot analysis, with Gapdh expression as the loading control. Insets: Representative western blots. (C) Nitrotyrosine levels using flow cytometry of the H9c2 cells. Data are the means ± SEM for the percentages of nitrotyrosine-positive cells, from at least 3 independent experiments, each performed in duplicate. (D) Effects of L-NAME on apoptosis induced under the same conditions. Data are the means ± SEM for the percentages of hypodiploid nuclei, from at least 3 independent experiments, each performed in duplicate. ^*P<0.05, ^**P<0.005, vs. control; °P<0.05 vs. cells treated with Doxo under the same experimental conditions; and ^#P<0.05 and ^##P<0.005, for cells co-treated with Doxo and L-NAME vs. Doxo (one-way ANOVA and multiple comparisons by Bonferroni's test). Doxo, doxorubicin; Rad, radicicol.