Nf κb p65 antibody

Paeoniflorin (> 98% purity) was obtained from Shanghai Daibo Chemical Technology Co., Ltd (Shanghai, China); HU308 (a selective CB₂R agonist) and AM630 (a selective CB₂R antagonist) were obtained from Cayman Chemicals (Ann Arbor, MI, USA). CB₂R antibody was obtained from Thermo Fisher Scientific Inc. (Waltham, MA, USA), NF-κB p65 antibody was obtained from Abcam Inc (Cambridge, MA, UK), Phospho-Akt (Ser473), Akt, mTOR, Phospho-mTOR (Ser2448) and Phospho-IκBα (Ser32) antibodies were obtained from Cell Signaling Technology Inc. (Beverly, MA, USA), iNOS, CD206, IκBα and lamin B1 antibodies were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA), CD68 antibody was obtained from AbD Serotec (Kidlington, Oxford, UK), Iba1 was from Wako Pure Chemical (Osaka, Tokyo, Japan), and phospho-PI3K p85α (Tyr467)/phospho-PI3K p55γ (Tyr199), PI3K and β-actin antibodies were obtained from Bioworld Technology Inc. (Minneapolis, MN, USA). An NO assay kit was obtained from Beyotime Institute of Biotechnology (Jiangsu, China).

BrMC was synthesized according to previous described 26 (link). The molecular formula is C₁₆H₁₁BrO₄, the character is yellow like powder, the molecular weight is 347, and its purity is 99.5% by HPLC. BrMC was diluted with dimethyl sulfoxide (DMSO) to 10 mmol/L, the desired concentration was diluted to by DMEM medium before application. TGF-β and TNF-α from Sino Biological (Beijing,China). NF-κBP65 inhibitors (SN50) and FoxM1 specific inhibitors, thiostrepton (THI), are purchased from Sigma-Aldrich (St. Louis, MO, USA). NF-κBp65 antibody was purchased from Abcam (Burlingame, CA, USA). The Rabbit anti human β-actin monoclonal antibody was derived from Proteintech (Chicago, USA). The following antibodies were purchased from CST (Massachusetts, USA): Anti FoxM1, CD44, Oct4, CD133 antibodies. The recombinant adenovirus containing pHBad-MCMV-GFP plasmids and pHBad-MCMV-GFP-NF-κBp65 plasmids (virus titer: 10¹⁰pfu/mL, 2 mL) was prepared by Han Heng Biotech Corp (Shanghai, China). The FoxM1expressing lentivirus, ENi.s (enhanced infection reagents and Polybrene were purchased from Genechem (Shanghai, China), puromycin purchased from Jikai gene company, Catalog No.REVG0002, Shanghai, China. Opti-MEM reagent is the product of Invitrogen (China, Shanghai).

Reagents used were CA125 IRMA kit (Tianjin Xiehe Medical Science & Technology Co., Ltd., cat. no. RC70417); E₂, P, FSH, and LH ELISA kits (Cayman Chemical, Ann Arbor, MI, USA, cat. no. 501890, 582601, 500710, and 500720); TLR4 antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA, cat. no. Sc-293072); NF-κB/P65 antibody (Abcam, Cambridge, UK, cat. no. AB16502); Beclin-1 antibody (ProteinTech, Wuhan, China, cat. no. 11306-1-AP); SQSTM1/P62 antibody (Bioss, Beijing, China, cat. no. Bs-55207r); GAPDH (Servicebio, Wuhan, China, cat. no. P04406); and HRP-labeled Goat Anti-Rabbit IgG (Servicebio, Wuhan, China, cat. no. GB23303).