Rabbit anti p ampkα thr172

For immunocytochemistry assays, primary astrocytes were fixed with 4% paraformaldehyde (PFA) for 15 min at room temperature. All the cells were blocked with 5% goat serum in 0.01 mM phosphate-buffered saline (PBS, pH 7.4) with 0.3% Triton-X-100 for 2 g h at room temperature and then incubated overnight at 4 °C with primary antibodies, including mouse anti-GlyT1 (1:25; Proteintech Group, Inc.), mouse anti-BDNF (1:50; Proteintech Group, Inc.), and rabbit anti-p-AMPKα (Thr172) (1:50; Affinity Biosciences, Inc.). Then, the cells were incubated with TRITC–conjugated goat anti-rabbit IgG or FITC–conjugated goat anti-mouse IgG (1:100; Proteintech Group, Inc.) for 2 h at room temperature. Finally, the cells were sealed with an antifluorescence attenuating tablet containing DAPI. Omission of the primary antibody served as a negative control. Images were captured by confocal laser scanning microscopy (FluoView FV 1000; Olympus, Tokyo, Japan). Image-Pro Plus 6.0 software was applied to quantify the fluorescence intensity of images, and GraphPad Prism 7.0 software was used to generate graphs.