Hela cell line

Dulbecco's modified Eagle's medium (DMEM), Roswell Park Memorial Institute (RPMI) medium, fetal bovine serum (FBS) and penicillin/streptomycin (P/S) were purchased from Welgene Inc. WST-1 and bovine serum albumin (BSA) were purchased from Sigma-Aldrich; Merck KGaA. A TUNEL assay kit was obtained from Promega Corporation. Hoechst dye, Mito-tracker^™ Red CMXRos and the Alexa-Fluor 488-conjugated donkey anti-Rabbit antibody were purchased from Thermo Fisher Scientific, Inc. The anti-Ki-67 antibody was purchased from Abcam (cat. no. ab15580). The aspirin conjugated DK143 [(E)-2-(4-(2,3-dimethoxyphenyl)acryloyl)-4-methoxyphenyl-2-acetoxybenzoate, AS-DK143] and AS-DK143-loaded methoxy poly(ethylene glycol)-poly(L-lactide) NPs (AS-DK143-NPs) were prepared and characterized according to our previous report (22 (link)). HeLa cell line, HepG2, which is a hepatoblastoma cell line, and AGS cell line were purchased from the Korean Cell Line Bank; Korean Cell Line Research Foundation. BXPC-3 cells were obtained from the Japanese Collection of Research Bioresources Cell Bank.

The HeLa cell line, obtained from Korean Cell Line Bank, were maintained in Dulbecco's Modified Eagle's medium supplemented with 10% fetal bovine serum (InvivoGen, San Diego, CA, USA), 100 U/ml penicillin-streptomycin (GenDepot, Katy, TX, USA) and 25 µg/ml normocin (InvivoGen). The HeLa cells were detached from dishes with trypsin-EDTA (GenDepot), and cell pellets were collected by centrifugation (400 × g for 5 min). The pellets were washed once with PBS and recovered by centrifugation (400 × g for 5 min). To obtain the cytosolic proteins, the cell pellet was lysed with lysis buffer (20 mM Tris, 20 mM NaCl, 1 mM EDTA, 5% glycerol, 0.1% Triton X-100 and 0.1% β-mercaptoethanol; pH 5.7; 4°C; 12 h) and the cell debris was removed by centrifugation (400 × g for 5 min).