P p65 s536

Ginsenoside panaxatriol was obtained from Must Bio-Technology (Chengdu, China). PTX was purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s Modified Eagle Medium (DMEM) (11995-040), F-12 nutrient mixture (Ham) and fetal bovine serum (FBS) were bought from Life Technologies (Grand Island, NY, USA). MammoCul medium (human) and supplements were purchased from STEMCELL Technologies (Vancouver, BC, Canada). CellTiter-Glo luminescent cell viability assay kits were purchased from Promega Corporation (Madison, WI, USA). iScript gDNA Clear cDNA Synthesis Kits and iTaq Universal SYBR Green Supermix Kits were purchased from Bio-Rad Laboratories (Hercules, CA, USA). p-IRAK1 S376, IRAK1, p-P65 S536, P65, p-ERK1/2, ERK1/2, BAX, BCL-2 and MCL-1 antibodies were supplied by Cell Signaling Technology (Danvers, MA, USA). Beta-actin antibody was purchased from Sigma–Aldrich (St. Louis, MO, USA).

For Western blotting assays, the following antibodies were used: A1CF (1:300, ORIGENE, Inc. Rockville, MD, USA, AP50047PU-N), NKRF (1:2000, Proteintech, Inc. Rosemont, IL, USA, 14693-1-AP), IFN-β (1:300, Proteintech, Inc., 27506-1-AP), p65 (1:1000, Cell Signaling Technology, Inc. Danvers, MA, USA, 6956), p50/105 (1:1000, Cell Signaling Technology, Inc., 13586), p-p65 (Ser536) (1:1000, Cell Signaling Technology, Inc., 3033), IκBα (1:1000, Cell Signaling Technology, Inc. 4814), p-IκBα (Ser32) (1:1000, Cell Signaling Technology, Inc., 2859), NPM1 (1:1000, Proteintech, Inc., 60096-1-Ig), β-actin (1:5000, Sigma-Aldrich, Inc. St. Louis, MO, USA, A1978), and GAPDH (1:5000, Sigma-Aldrich, Inc., SAB2701825).
For Immunofluorescence and immunohistochemistry, the following antibodies were used: A1CF (1:50, ORIGENE, Inc., AP50047PU-N), NKRF (1:200, Santa Cruz Biotechnology, Inc. Santa Cruz, CA, USA, sc-365568), IFN-β (1:50, Proteintech, Inc., 27506-1-AP), p65 (1:200, Cell Signaling Technology, Inc., 6956), and p-p65(S536) (1:200, Cell Signaling Technology, Inc., 3033).

Antibodies against mTOR (1:1000), p-mTOR (Ser2448) (1:1000), p-ULK1 (S757) (1:1000), Akt (1:1000), p-Akt (S473) (1:1000), NF-κB(p65) (1:1000), p-p65 (S536) (1:1000), acetylated lysine (Ace-lys) (1:1000), and Beclin1(1:1000) were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA). Antibodies targeting Col II (1:1000 for western blotting analysis, 1:200 for immunohistochemistry staining), ULK1 (1:1000), and p62 (1:1000) were purchased from Abcam Inc. (Cambridge, MA, USA). Antibodies against Aggrecan (1:1000 for western blotting analysis, 1:200 for immunohistochemistry staining) and β-actin (1:40,000) were purchased from Sigma-Aldrich in China (Shanghai, China). Rabbit IgG and Protein A/G PLUS-Agarose were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against LC3B (1:1000 for western blotting analysis, 1:200 for immunohistochemistry staining), Lamin B1 (1:1000), and β-tubulin (1:1000) were purchased from Novus Biologicals, Inc. (Littleton, CO, USA), Protein Tech Group (Chicago, IL, USA), and Sangon Biotech (Shanghai, China), respectively. Other reagents were of the highest grade commercially available.