Dulbecco s modified eagle medium dmem

Dulbecco's Modified Eagle Medium (DMEM) and fetal bovine serum (FBS) were purchased from Cytiva (Marlborough, MA, USA). LPS-PG was obtained from Invivogen (San Diego, CA, USA). MG-132, LY294002, U0126, and CGA were purchased from Sigma-Aldrich (St. Louis, MI, USA) which were dissolved in dimethyl sulfoxide (DMSO).

Human hepatocarcinoma cell lines that lacked TM4SF5 expression (SNU449 and empty-vector control SNU449C_p cells) or that expressed wild-type (WT) TM4SF5 [(exogenous expression) SNU449T₇, (endogenous expression) HepG2, Huh7, and HepG2 cells] were used for in vitro experiments and were previously described [17 (link)]. Cells were purchased from either the Korean Cell Bank (Seoul, Korea) or American Type Culture Collection (ATCC, Manassas, VA, USA). Hepatocytes were cultured in Roswell Park Memorial Institute (RPMI) 1640 Medium or Dulbecco’s Modified Eagle Medium (DMEM) (Cytiva, Marlborough, MA, USA) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (GenDEPOT Inc., Barker, TX, USA) at 37 °C in 5% CO₂. Cells were stably infected with empty pLJM1-EGFP (Addgene) or TM4SF5-encoding pLJM1-EGFP lentiviral vectors. Human NK92 cells were cultured in α-Minimum Essential Medium (MEM) (Invitrogen, Grand Island, NY, USA) containing 200 U/mL recombinant human interleukin (IL)-2 (PeproTech, Rocky Hill, NJ, USA), 12.5% FBS, and 12.5% fetal horse serum (GenDEPOT Inc.). Stable cells were cultured in RPMI 1640 (WelGene) containing 10% FBS, geneticin (G418; 250 μg/mL), and antibiotics (Invitrogen, Grand Island, NY, USA). Cells were passaged every 3–4 days at ratios specified by the suppliers. Cells were monitored for mycoplasma contamination.

Antibodies against α-SMA, collagen type I, fibronectin, E-cadherin, vimentin, phospho-Smad3, phospho-p38, p-38, phospho-ERK1/2 (T202/Y204), and ERK1/2 were obtained from Cell Signaling Technology (Beverly, MA). Antibodies against LC3II and p-62 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Anti-mouse immunoglobulin G and anti-rabbit immunoglobulin G fluorescent-conjugated secondary antibodies obtained were from LI-COR Biotechnology (Nebraska, United States). Dulbecco’s Modified Eagle Medium (DMEM) was obtained from Cytiva, United States. Fetal bovine serum (FBS) was obtained from Grand Island Biological Company (Gibco, United States). DAPI was obtained from Yeasen Biotech Co., Ltd. (Yeasen Shanghai). SB-431542 was obtained from MedChemExpress (MCE, United States). Piceatannol was from Chengdu Herbpurify Co., Ltd.

The human cancer cell lines (H1299 (Cat No. FH0908), A549 (Cat No. FH0045), H292 (Cat No. FH0085) obtained from FuHeng Biology (Shanghai, China); SW900 (Cat No. BNCC275987) obtained from BNBIO (Beijing, China); PC9 (Cat No. MZ‐2682) obtained from Mingzhou Bio), were cultured as previously reported.17 (link), 18 (link) The human bronchial epithelial cell lines (HBE; BNBIO, Cat No. BNCC338600) and BEAS‐2b (FuHeng Biology, Cat No. FH0319) were cultured in Dulbecco's modified eagle medium (DMEM) purchased from HyClone Laboratories Inc. supplemented with 5% foetal bovine serum and 1% penicillin/streptomycin solution, at 37°C in a humidified atmosphere containing 5% CO₂.

Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), and penicillin-streptomycin (PS) were purchased from Hyclone Laboratories (Logan, UT, USA). Aβ_25–35 peptide, dimethylsulfoxide anhydrous (DMSO), mouse anti-SYN, triton X-100, slide mounting medium for histology, paraformaldehyde (PFA), 3,3-diaminobenzidine (DAB), DPPH, ABTS, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA), phosphate buffer (PB), and phosphate buffered saline (PBS, pH 7.4) were purchased from Sigma Aldrich (St. Louis, MO, USA). Rabbit anti-PSD-95 and goat anti-NQO1 were purchased from Abcam (Cambridge, UK). Rabbit anti-HO-1 was purchased from Enzo Life Sciences (Farmingdale, NY, USA). Rabbit anti-phosphorylated CREB (pCREB) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Mouse anti-NeuN was purchased from Millipore (Billerica, MA, USA). Normal goat serum (NGS), avidin-viotin complex (ABC), and biotinylated anti-mouse and anti-rabbit antibodies were purchased from Vector Lab (Burlingame, CA, USA).

Cartilage tissues were collected from the knees of 10 OA patients diagnosed by the ACR criteria at the time of surgery (Table S1). After written informed consent was acquired, the human knee OA cartilage was obtained from surgical waste of patient undergoing TKA. The cartilage was rinsed with phosphate buffered saline (PBS) containing the following: 137 mM NaCl; 2.7 mM KCl; 10 mM Na₂HPO₄, and 1.8 mM KH₂PO₄ and minced into fine pieces. Enzymatic digestion with pronase (Sigma-Aldrich, St. Louis, MO, USA) and collagenase type II (Worthington Biochemical Corp., Lakewood, NJ, USA) and collagenase type II at temperature of 37 °C for 2 h was used to isolate chondrocytes from the articular cartilage. The digested articular cartilage was centrifuged at 1000× g rpm for 5 min, and supernatant was then discarded. Afterwards, the pellet was resuspended, seeded, and cultured in chondrogenic growth media (Dulbecco’s Modified Eagle Medium (DMEM; HyClone Laboratories, South Logan, UT, USA)/F12, 10% FBS (HyClone Laboratories, South Logan, UT, USA), 1% glutamax (GIBCO—BRL, Grand Island, NY, USA), 1% antibiotic-antimycotic (GIBCO—BRL, Grand Island, NY, USA), 0.05 mg/mL of ascorbic acid (Sigma-Aldrich, St. Louis, MO, USA). Culture media was replaced every 2–3 days, and knee OA chondrocyte passages 1–3 were utilized in the subsequent experiments.