Cell counting kit 8

DSF was purchased from Selleckchem (Houston, TX, USA). TR, TMTM, Sodium diethyldithiocarbamate trihydrate, and MMS were purchased from Sigma-Aldrich (St. Louis, MO, USA). AHA and mitomycin were purchased from Wako Pure Chemical Industries (Osaka, Japan). ILM was purchased from Focus Biomolecules (Plymouth Meeting, PA, USA). DETC-MeSO and DDTC-MeSO were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The FDA-Approved Drug Screen-well Library and Cell Counting Kit (CCK)-8 were obtained from Enzo Life Sciences (Farmingdale, NY, USA) and Dojindo (Kumamoto, Japan), respectively. PLC/PRF/5 cells were authenticated by the short tandem repeat method (Bex, Tokyo, Japan) in 2016 and cultured in Dulbecco’s modified Eagle medium containing fetal bovine serum according to the protocol (www.atcc.org/Products/All/CRL-8024.aspx#culturemethod) of American Type Culture Collection (ATCC) (Manassas, VA, USA). NK92MI cells were also obtained from ATCC and cultured in Alpha Minimum Essential medium containing horse and fetal bovine serums following the protocol (www.atcc.org/Products/All/CRL-2408.aspx#culturemethod). PXB cells were purchased from Phoenix Bio (Hiroshima, Japan). Cells were cultured at 37°C and 5 % CO₂.

Polyclonal antibodies against collagen III (catalog ab83829), NT3 (catalog ab53685), and the NT3-neutralizing antibody (catalog ab6203) were purchased from Abcam (Cambridge, MA, USA). Recombinant human NT3 was obtained from US Biological (San Antonio, TX, USA). The TLR4-neutralizing antibody (catalog HTA125) was purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The thymidine analog 5-bromo-2′-deoxyuridine (BrdU) cell proliferation kit and antibodies against phosphorylated Akt (catalog 9271), total Akt (catalog 9272), phosphorylated ERK1/2 (catalog 9010), and total ERK1/2 (catalog 4695), MMP-9 (catalog 3852), and Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (catalog 2118, clone number 14C10) were purchased from Cell Signaling, Inc. (Beverly, MA, USA). The Trk inhibitor K252a was obtained from Calbiochem (Darmstadt, Germany). The Akt inhibitor MK2206 and ERK1/2 inhibitor PD98059 were purchased from Selleckchem (Houston, TX, USA). The Cell Counting Kit (CCK)-8 was purchased from Enzo Life Sciences International (Plymouth Meeting, PA, USA). Medium 199 was purchased from Lonza (Walkersville, MD, USA). Picro-Sirius Red and other reagents were purchased from Sigma-Aldrich Chemical (St Louis, MO, USA).

Anti-phospho-AMPK, phospho-p38, phospho-ERK, phospho-JNK, phospho-Akt, AMPK, poly (ADP-ribose) polymerase (PARP), caspase-3, caspase-9, Bcl-2, Bcl-extra-large (Bcl-xL), and Bcl-2-associated X protein (Bax) antibodies (Cell Signaling, Danvers, MA, United States). Anti-p38, ERK, JNK, Akt, γH2AX, β-actin, and α-tubulin antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, United States). SB203580 was obtained from Sigma-Aldrich (St. Louis, MO, United States). Compound C (CC) was purchased from MedChemExpress (Monmouth Junction, NJ, United States). Matrigel was obtained from BD Biosciences (San Diego, CA, United States). The cell counting kit (CCK)-8 was purchased from Enzo Life Sciences (Farmingdale, NY, United States). The mouse CRC cell line CT26 and MC38, human CRC cell line HT29 and SW620, and normal CCD-18co colon cell line were purchased from Korean Cell Line Bank (Seoul, South Korea) and maintained in Dulbecco’s modified Eagle’s medium (DMEM) and Roswell Park Memorial Institute (RPMI) 1640 at 37°C in a 5% CO₂ incubator.