Fig 2: Effect of decreased NKA α2 in the PVN on the level of NKA α2, PKC γ, and p-Rac1 in salt-induced hypertension. After microinjection of NKA α2 shRNA in the PVN, the protein expression of NKA α2, Protein Kinase Cγ (PKC γ) and phosphorylated Ras-related C3 botulinum toxin 1 (p-Rac1) in the PVN in different groups. (A) Immunohistochemistry for NKA α2 in the PVN in different groups. (B) Column diagram on the number of NKA α2 positive neurons in the PVN in different groups. (C) A representative immunoblot of protein expression and (D) densitometric analysis of NKA α2, PKC γ, and p-Rac1 in the PVN in different groups. (E) A representative immunoblot and (F) densitometric analysis of p-Rac1 in the PVN in different groups. * p < 0.05 versus the normal salt groups (the normal salt + NKA α2 shRNA or normal salt + scrambled shRNA group); # p < 0.05, the high salt + NKA α2 shRNA group versus the high salt + scrambled shRNA group. Values are expressed as the mean ± SEM (n = 6).

Fig 3: Effect of decreased NKA α2 expression in the PVN on the oxidative stress in salt-induced hypertension. Rac1 is one of the important NAD(P)H subunits, which contributes to oxidative reactive oxygen species (ROS) production. DHE can detect the superoxide anion levels in the tissues. (A) Immunofluorescence for DHE detecting. (B) Column diagram showing the immunofluorescent intensity of DHE in the PVN in different groups. ELISA results showing the levels of MDA, SOD, GSH, GSSH, GSH/GSSH, and CAT in the PVN. (C) The level of MDA, (D) SOD activity, (E) GSH, (F) GSSH, (G) GSH/GSSH ratio, and (H) CAT activity in the PVN in the different groups. * p < 0.05 versus the normal salt groups (the normal salt + NKA α2 shRNA or normal salt + scrambled shRNA group); # p < 0.05, the high salt + NKA α2 shRNA group versus the high salt + scrambled shRNA group. Values are expressed as the mean ± SEM (n = 6).