Jurkat t lymphocyte and hek 293t kidney cells

Jurkat T-lymphocyte and HEK 293T kidney cells (American Type Culture Collection) with 5 × 10 5 cells/mL in 6-well plate, were transfected with LV-CRISPRi-lincRNA-p21 or LV-CRISPRi-GFP for 48 hr in the presence of polybrene (8 µg/ml, Sigma-Aldrich), and were incubated with 5 µg/ml and 0.5 µg/ml of puromycin, respectively [26] . The puromycin selection process was up to one month in order to select successfully transduced stable transfectants con rmed by qRT-PCR analyses. Doxorubicin (Dox)-induced cell apoptosis HEK 293T, HK-2, Jurkat cells or transfectants were seeded with 1 10 6 cells/mL in 6-well plate in the presence of Dox (TTY Biopharm) for 24 hr under 37 ℃, 5 % CO 2 incubation [11] . These cells were further stained with Annexin V and 7-AAD (BD Pharmingen) to detect apoptotic and dead cells, respectively, by ow cytometric analyses. Apoptotic cells were de ned as Annexin V+ and 7-AAD-in this study.