Figure 8.
β-adrenergic cardiac stress evokes S-nitrosylation lateralized Cx43 proteins. (A) Representative images of analysis performed by PLA of the interaction between Cx43 and S-nitrosylation. Plasma membrane stained with wheat germ agglutinin (WGA) and S-nitrosylated Cx43 (Cx43-SNO) are shown in green and red, respectively. Scale bar: 50 μm. (B) Quantification of PLA dots signals observed in A. P = 0.015 WT vs. WT + Iso; P = 0.0001 WT + Iso vs. S3A + Iso; P = 0.0001 S3A vs. S3A + Iso. Comparisons between groups were made using two-way ANOVA plus Tukey’s post-hoc test. The number in parentheses indicates biological replicate. (C) The relative percentage of Cx43-NO dots signal at the lateral side of cardiomyocytes detected in A. Note that S3A-Iso enhanced lateralized signal. P = 0.0001 WT vs. S3A + Iso; P = 0.0001 WT + Iso vs. S3A + Iso; P = 0.0001 S3A vs. S3A + Iso. Comparisons between groups were made using two-way ANOVA plus Tukey’s post-hoc test. The number in parentheses indicates biological replicate. (D) Analysis of cell areas are detected in WT and S3A mice under a vehicle and upon Iso stimulation. (E) Top and middle gels were loaded with S-nitrosylated proteins pulled down using the biotin switch assay. The top gel was then blotted against Cx43. The middle gel is the corresponding ponceau staining. The lower blot was loaded using total cardiac proteins and blotted against Cx43 from heart samples blotted against Cx43. The bottom graph is the quantification for two independent blots using the ratio for SNO-Cx43/total Cx43. P = 0.0270 WT vs. WT + Iso; P = 0.0074 WT + Iso vs. S3A + Iso; P = 0.0002 S3A vs. S3A + Iso. Comparisons between groups were made using two-way ANOVA plus Tukey’s post-hoc test. The number in parentheses indicates biological replicate. Source data are available for this figure: SourceData F8.

β-adrenergic cardiac stress evokes S-nitrosylation lateralized Cx43 proteins. (A) Representative images of analysis performed by PLA of the interaction between Cx43 and S-nitrosylation. Plasma membrane stained with wheat germ agglutinin (WGA) and S-nitrosylated Cx43 (Cx43-SNO) are shown in green and red, respectively. Scale bar: 50 μm. (B) Quantification of PLA dots signals observed in A. P = 0.015 WT vs. WT + Iso; P = 0.0001 WT + Iso vs. S3A + Iso; P = 0.0001 S3A vs. S3A + Iso. Comparisons between groups were made using two-way ANOVA plus Tukey’s post-hoc test. The number in parentheses indicates biological replicate. (C) The relative percentage of Cx43-NO dots signal at the lateral side of cardiomyocytes detected in A. Note that S3A-Iso enhanced lateralized signal. P = 0.0001 WT vs. S3A + Iso; P = 0.0001 WT + Iso vs. S3A + Iso; P = 0.0001 S3A vs. S3A + Iso. Comparisons between groups were made using two-way ANOVA plus Tukey’s post-hoc test. The number in parentheses indicates biological replicate. (D) Analysis of cell areas are detected in WT and S3A mice under a vehicle and upon Iso stimulation. (E) Top and middle gels were loaded with S-nitrosylated proteins pulled down using the biotin switch assay. The top gel was then blotted against Cx43. The middle gel is the corresponding ponceau staining. The lower blot was loaded using total cardiac proteins and blotted against Cx43 from heart samples blotted against Cx43. The bottom graph is the quantification for two independent blots using the ratio for SNO-Cx43/total Cx43. P = 0.0270 WT vs. WT + Iso; P = 0.0074 WT + Iso vs. S3A + Iso; P = 0.0002 S3A vs. S3A + Iso. Comparisons between groups were made using two-way ANOVA plus Tukey’s post-hoc test. The number in parentheses indicates biological replicate. Source data are available for this figure: SourceData F8.

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