Sw1900

Human liver cancer cell lines HUH7, Hep3B, PLC/PRF/5, SK-Hep1, SNU-449, HepG2, pancreatic cancer cell line SW1900 and PANC1, lung cancer cell line H69 and A549, breast cancer cell line MDA231, and cervical cancer cell line Hela were obtained from the American Type Culture Collection (ATCC, USA) and cultured with Dulbecco’s Modified Eagle Medium (DMEM, Gibco, Thermo Fisher, USA). All cultured media were supplemented with 10% fetal bovine serum (FBS) and 1% penicillin–streptomycin (10,000 Units/mL of penicillin and 10000 µg/mL of streptomycin, Life Technologies, USA). All cell lines were in the incubator with 5% CO₂ at 37 ℃ (CO₂ water-jacketed incubator, Nuaire®, USA). All cells were identified by short-tandem-repeat profiling and without mycoplasma contamination.

The human pancreatic cancer cell lines Aspc‐1, Bxpc‐3, CFPAC‐1, MiaPaCa‐2, PANC‐1 and SW1900 were purchased from American Type Culture Collection (ATCC, Rockville, MD, USA). MiaPaCa‐2, PANC‐1 were maintained in DMEM supplemented with 10% foetal bovine serum, 100 units/ml penicillin and 100 mg/ml streptomycin in a humidified incubator containing 5% CO₂ in air at 37°C. Aspc‐1, Bxpc‐3, CFPAC‐1 and SW1900 were maintained in RPMI 1640 containing supplements as above. For all experiments, the cell lines were treated in serum‐free medium (0.1% bovine serum albumin, 5 mg/ml transferrin and 5 ng/ml sodium selenite and antibiotics). Hypoxic conditions (1% O₂) were created with a three‐gas incubator MiniGalaxy A (RS Biotech, Irvine, UK) by injection of N2 (1% O₂/94% N₂/5% CO₂ atmosphere, 37°C). In other experiments hypoxia mimetic conditions were chemically generated by treating cells with 100 mM cobalt chloride (CoCl₂; Sigma‐Aldrich, St. Louis, MO, USA) for the indicated times.

Four human PC cell lines (SW1900, PANC-1, AsPC-1, and BxPC-3) and a normal human pancreatic ductal epithelial cell line HPDE6-C7, were obtained from the American Type Culture Collection (Rockville, MD, USA). PC cell lines were routinely maintained in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum (FBS; Gibco, Grand Island, NY, USA) at 37 °C in a humidified incubator set at 5% CO₂. HPDE6-C7 cells were cultured in keratinocyte serum-free medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% FBS and antibiotics in a humidified incubator set at 37 °C and 5% CO₂ atmosphere. The primary antibody against TRIM29 (Abcam, ab108627) was used (1:1,000 dilution) for western blotting and for immunohistochemistry (1:200 dilution). The primary antibody against YAP1 (Abcam, ab205270) was used (1:1,000 dilution) for western blotting and for immunohistochemistry (1:150 dilution). The primary antibodies against Ub (Abcam, ab134953), PCNA (Abcam, ab92552), Cyclin D1 (Abcam, ab40754), BCL2 (Abcam, ab32124), Bax (Abcam, ab32503), and Caspase 3 (Abcam, ab32351) were used at 1:1,000 dilution for the western blot, and the primary antibody against GAPDH (Abcam, ab181602) was used at a 1:3,000 dilution for western blotting (Additional files 1, 2 and 3).