Fig. 2

Immunofluorescence analysis showing HYP increased nNOS expression in primary hippocampal neurons. Cultured hippocampal neuronal cells were treated with 10 nM E2 and 0.1 or 1 µM HYP for 24 h. Cells were incubated with primary nNOS antibody (red) and a neuronal marker neurofilament (NF, green). Cells were then incubated with secondary antibody, and nuclei were stained with DAPI (blue). Confocal microscopy captured immunofluorescence images, and nNOS intensity was quantified. The merged image shows the nNOS-positive cells. Results are presented as mean ± SE. *P < 0.05, **P < 0.01, ***P < 0.001 vs. CTL (vehicle-only treated cell). E2, 17β-estradiol; HYP, hyperoside; nNOS, neuronal nitric oxide synthase; NO, nitric oxide