Figure 5.

The regulation of eNOS activity by nicotine microinjected into the NTS. (A) The expression of NeuN (a) and eNOS (b) in the NTS of WKY rats as determined by immunofluorescent staining. (c) The merged image of NeuN-FITC (green, panel a) and eNOS-Rhod (red, panel c). (d) The merged image of NeuN-FITC, eNOS-Rhod and DAPI. The original magnification was ×400. Scale bar = 100 µm. (B) The immunoblot depicts the phosphorylation of eNOS^S1177 proteins in the NTS without (lane 1) or with (lane 2) nicotine treatment. Phosphorylated eNOS^S1177 was not increased in the nicotine-microinjected group as compared with the control group. (C) The immunoblot depicts the phosphorylation of ERK1/2^T202/Y204 proteins in the NTS without (lane 1) or with (lane 2) nicotine treatment. Phosphorylated ERK1/2^T202/Y204 was not increased in the nicotine-microinjected group compared with the control group. (D) The immunoblot depicts the phosphorylation of Akt^S473 proteins in the NTS without (lane 1) or with (lane 2) nicotine treatment. (E) Lysates from NTS were assayed and used for co-immunoprecipitation of calmodulin and eNOS. Total lysates were also analysed by immunoblot with antibodies to eNOS and calmodulin. eNOS interacts with calmodulin. Values are shown as mean ± SEM, n = 6. P > 0.05 versus control group. BP, blood pressure; DAPI, 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride; eNOS, endothelial nitric oxide synthase; ERK, extracellular signal-regulated kinases; FITC, fluorescein isothiocyanate; HR, heart rate; NeuN, neuronal nuclei; nNOS, neuronal nitric oxide synthase; NTS, nucleus tractus solitarii.