Hek293

Human embryonic kidney 293 (HEK293) (KAC Co, Ltd (Kyoto, Japan)) and HEK293T cells were cultured in Eagle’s minimum essential medium (FUJIFILM Wako Pure Chemical, Osaka, Japan) supplemented with 1 × nonessential amino acids (FUJIFILM Wako Pure Chemical) and 10% (v/v) fetal bovine serum (FBS). HEK293 cells were seeded at a density of 7.5 × 10⁵ cells/well in a 24-well plate and incubated with or without IL-1β (1 ng/ml) and TGF-β1 (10 ng/ml) (FUJIFILM Wako Pure Chemical).
Normal human dermal fibroblasts (NHDFs), Detroit 551, were obtained from the American Type Culture Collection (VA, USA). The Detroit 551 cells were maintained in Eagle’s minimum essential medium supplemented with 1 × nonessential amino acids, 1 mM sodium pyruvate, and 10% (v/v) FBS. The Detroit 551 cells were seeded at a density of 2.5 × 10⁴ cells/well in a 24-well plate and incubated with or without a mixture of the cytokines (0.1–1.0 ng/ml, TNF-α, IL-1β, and IL-6), IL-1β (1.0 ng/ml) or TGF-β (10 ng/ml) (FUJIFILM Wako Pure Chemical).

As the CR-1 overexpressing cells, human breast carcinoma cell line T47D [18 (link),19 (link)] and human teratocarcinoma cell line N-tera2 D1 [20 (link),21 (link)] were obtained from American Type Culture Collection (ATCC) (Manassas, VA, USA), and human colon cancer cell line GEO [22 (link),23 (link)] was kindly provided by Dr. David Salomon under the MTA of National Cancer Institute, Bethesda, MD. HEK293 cells [24 (link),25 (link)] were obtained from ATCC. ExpiCHO-S cells were from Thermo Fisher Scientific (Waltham, MA, USA). T47D, GEO and HEK293 cells were maintained in DMEM with high glucose formula (WAKO, Tokyo, Japan) containing 10% FBS (Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin–streptomycin (WAKO, Tokyo, Japan) at 37 °C under the atmosphere of 5% CO₂. ExpiCHO-S cells were cultured in ExpiCHO Expression Medium (Thermo Fisher scientific, Waltham, MA, USA) at 37 °C under 8% CO₂ shaking at 120 rpm [26 (link),27 (link)].