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. 2021 Jan 4;12(1):4. doi: 10.1038/s41419-020-03338-4

Fig. 4. eNOS contributes to NO production and S-nitrosylation of selected proteins.

Fig. 4

A Relative increase of NO after the addition of 200 ng/ml BDNF to cortical cell cultures previously transfected with an shRNA targeting eNOS, sh-Luc shRNA, or not transfected controls. B Mean slopes of NO production are shown in n = 4–6 independent experiments, *p < 0.5 by two-way ANOVA followed by Bonferroni post-test. C The biotin-switch assay was used to pull down S-nitrosylated proteins. Western blots detecting p65 subunit and tubulin 1α in the pull-downs of cortical and hippocampal cultures are shown. Cell cultures were transfected with shRNA targeting eNOS or scrambled shRNA. D Densitometric quantification of the S-nitrosylated (SNO) levels of NF-κB subunit p65 and tubulin α-1A. Results obtained from n = 4–6 independent experiments. *p < 0.05; **p < 0.01; ***p < 0.001 by two-tailed t-test. CX cortical cultures, HP hippocampal cultures, Control not transfected cortical neurons. Sc scrambled shRNA sequence, eNOS short interfering RNA against eNOS, C is a negative control for the biotin switch assay (pull-down of samples in which reduction with ascorbate was omitted).