β actin bs 10966r

The total DNA/RNA/Protein Kit (R6734, Omega Bio-tek, Norcross, GA, USA) was utilized to extract total protein from hippocampal tissue according to the manufacturer’s instructions. Western blot was conducted for CREB (#9197, Cell signaling technology, USA), p-CREB (#9198, Cell signaling technology, USA), BDNF (ab108319, Abcam, UK), AKT (#9272, Cell signaling technology, USA), p-AKT (#4060, Cell signaling technology, USA), PSD95 (#3450, Cell signaling technology, USA), GABAA (GB113973, Servicebio, Wuhan, China), Synapsin-1 (SYN, #5297, Cell signaling technology, USA), Bax (#2772, Cell signaling technology, USA), Bcl-2 (#3498, Cell signaling technology, USA), Caspase-3 (#9662, Cell signaling technology, USA), Cleaved-caspase 3 (#9661, Cell signaling technology), NF-kB (P65) (#8242, Cell signaling technology, USA), NLRP3 (sc-134306, Santa Cruz, CA, USA), Caspase 9 (#9504, Cell signaling technology), Caspase 1 (#3866, Cell signaling technology), β-actin (bs-10966R, Bioss, Beijing, China) of hippocampal total protein. The density of protein bands was assessed using a computing densitometer. IL-1β was measured using ELISA kits (Nanjingjiancheng, Nanjing, China) according to the manufacturer’s protocols.

The phosphofructokinase 1 (PFK1) (sc-31711) antibody was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), while the phosphofructokinase 2 (PFK2) (bs-3528R), carnitine palmitoyltransferase-1A (CPT-1A) (bs-2047R), AMP-activated protein kinase (AMPK) (bs-1115R), β-actin (bs-10966R), phosphorylated PFK2 at Ser467 (bs-3331R), and phosphorylated pyruvate kinase M2 (p-PKM2) at Tyr105 (bs-3334R) antibodies were all purchased from Bioss Biotechnology (Beijing, China). The antibodies against PKM2 (15822–1-AP), hexokinase 2 (HK2) (22029–1-AP), α-ketoglutarate dehydrogenase (OGDH) (15212–1-AP), citrate synthase (CS) (16131–1-AP), isocitrate dehydrogenase 2 (IDH2) (15932–1-AP), lactate dehydrogenase B (LDHB) (14824–1-AP), and pyruvate dehydrogenase E1(PDHE1) (18068–1-AP) were all purchased from Proteintech Group (Chicago, IL, USA). The phosphorylated AMPK (Tyr172, 2531) antibody was acquired from Cell Signaling Technology (Beverly, MA, USA). The macrophage migration inhibitory factor (MIF) antibody was produced in our laboratory. Hematoxylin (AR11180–1) and eosin (AR11180–2) staining solutions were purchased from Boster Bioengineering Co. Ltd. (Wuhan, China), while the RIPA protein buffer and BCA protein assay kit (P0010) were procured from Beyotime Biotechnology (Shanghai, China). ECL-plus reagent (AR1111) was obtained from Bioss Biotechnology.