Rabbit anti hmox 1 antibody

Immunoblotting was conducted as described previously [34 (link)]. Briefly, cell lysates extracted using the Pierce M-PER Mammalian Cell Lysis Buffer supplemented with 1% SIGMAFAST^™ Protease Inhibitor Cocktail (Sigma-Aldrich, St. Louis, MO, USA) were denatured, separated on a 4–12% NuPage^® Novex^® Bis-Tris gradient gel (Life Technologies, Carlsbad, CA, USA) at 200 V for 45 min, and transferred onto a nitrocellulose membrane (LI-COR, Lincoln, NE, USA) at 30 V for 1 h. Proteins were detected by incubating first with primary antibodies (rabbit anti-HMOX-1 antibody, Cell Signaling, Danvers, MA, USA; mouse anti-acetylated α-tubulin, mouse anti-AKR1B10, rabbit-anti-DNAI1, and mouse anti-CDC20B, Sigma–Aldrich; mouse anti-CK13 and goat anti-MGMT, Santa Cruz, Dallas, TX, USA; mouse anti-involucrin, Thermo Fisher Scientific, Waltham, MA, USA; rabbit anti-ADH1C and rabbit anti-FANCD2, Abcam, Cambridge, MA, USA), followed by a 30-min incubation with the IRDye-conjugated secondary antibodies (LI-COR). Images were taken using an Odyssey^® CLx Imaging System (LI-COR). Densitometry was conducted using LI-COR Image Studio software.

Expression of key protein markers was measured at T1, T12, and PT20 using immunoblotting as described previously (Xiong et al. 2018 (link)). Briefly, proteins were denatured in a loading buffer containing 50 mM dithiothreitol (DTT), separated on a NuPage^® Novex^® 4–12% Bis–Tris gradient gel (Life Technologies, Carlsbad, CA), and transferred onto a nitrocellulose membrane (LI-COR, Lincoln, NE). Proteins were detected by incubating first with primary antibodies (rabbit anti-HMOX-1 antibody, Cell Signaling, Danvers, MA; mouse anti-acetylated α-tubulin, Sigma-Aldrich, St. Louis, MO; mouse anti-CK6, Santa Cruz, Dallas, TX; or mouse anti-involucrin, Thermo Fisher Scientific, Waltham, MA) and then IRDye-conjugated secondary antibodies (LI-COR). Images were taken using the LI-COR Odyssey^® CLx Imaging System and densitometry performed using the LI-COR Image Studio software.

The XYW (Tai Ji, China, batch number 1707029) and fluoxetine hydrochloride (FLX) (Patheon, France, 7686 A) were dissolved in pure water solution preparation (SOD, batch number 20180,309), malondialdehyde (MDA, batch number 20180,313) (GSH, batch number 20180,621), and GSH/glutathione disulfide (GSH/GSSG, batch number 20180,726) levels were assessed using commercially available kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Nitric oxide (NO) assay kit (Biyuntian, 0,72117,171,110, China). Rat interleukin 6 (IL6) ELISA Kit (MULTI SCIENCES, A30680231), rat interleukin 1β (IL1B) ELISA Kit (ExCell Bio, 21H183), rabbit anti-NTRK2 antibody (Cell Signaling Technology; Cat. No. #4603), rabbit anti-AKT1 antibody (Cell Signaling Technology; Cat. No. #4691S), rabbit anti-p-AKT1 antibody (Cell Signaling Technology; Cat. No. #4060S), rabbit anti-HMOX1 antibody (Cell Signaling Technology; Cat. No. #S2206S), rabbit anti-PIK3CA antibody (Servicebio; Cat. No. #LS190932), rabbit anti-GAPDH antibody (Servicebio; Cat. No. #GB11002), rabbit anti-SOD1 antibody (Novus Biologicals; Cat. No. #03253462C2B), rabbit anti-NFE2L2 antibody (Invitrogen; Cat. No. #TK2668681A), rabbit anti-brain-derived neurotrophic factor (BDNF) antibody (Abcam; Cat. No. #ab108319) and anti-rabbit IgG HRP-linked antibody (Servicebio; Cat. No. #GB23303).