Hle b3

Human lens epithelium B3 (HLE-B3) and 239T cell lines were obtained from American Type Culture Collection (ATCC; Rockville, MD) and cultured in Dulbecco’s modified Eagle’s medium (DMEM; Sigma-Aldrich) with 10 % (v/v) fetal bovine serum (FBS; Sigma) in a humidified atmosphere of 5 % CO₂ at 37 °C. We used the 293T cell line because this cell line is a common model for vector transfection with high transfection efficiency. We used the HLE-B3 for other experiments because the cells are considered a possibility host for intervention and UV radiation experiment [57 (link)]. HLE-B3 was cultured in six-well plates in DMEM without FBS for 24 h prior to treatment. Then, the HLE-B3 were treated with 10 μM 5-aza-dC (Sigma, St. Louis, MO) for 48 h and 5 μM MS275 (Selleckchem, Houston, TX, USA) for 12 h, and exposed to UVB light for 20 min. We harvested the cells on different time point (1000 J/m², XX-15B, Spectroline, Westbury, NY, USA), respectively. The intensity and dose of UBV were measured using a UVX Radiometer connected to a UVX-31 Sensor (both were from UVP Inc., San Gabriel, CA, USA). After exposure, the DMEM was immediately replaced by DMEM with 10 % FBS. At different time point, the cells were harvested for DNA, mRNA, and protein extraction.

Human lens epithelial cells (HLE-B3; American Type Culture Collection, Manassas, VA, USA) were grown in Dulbecco’s Modified Eagle Medium (Thermo Fisher Scientific Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich; St. Louis, MO, USA), 100 U/ml penicillin (Duchefa; RV Haarlem, the Netherlands), and 100 μg/ml streptomycin (Duchefa). HLE-B3 cells were infected with shBRD4 (Sigma Aldrich) viral particles for 24 h. All cultured cells were incubated at 37°C in a humidified atmosphere containing 5% CO₂.