Bcl2l11 overexpressing lentivirus

HEK-293T cells were obtained from the American Type Culture Collection. The Tsc2 -/-MEF cell line, which has been described previously, was a gift from Hongbing Zhang (Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, People's Republic of China) [12, 13] . All cell lines were cultured in DMEM supplemented with 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 mg/ml of streptomycin at 37 °C in a humidified 5% CO 2 atmosphere.
HEK-293T or Tsc2 -/-MEF cells were seeded in 6-well plates, and transfection was conducted after 24 h. The BCL2L11-overexpressing lentivirus and the control lentivirus were purchased from GenePharma (Shanghai, China); 10 6 recombinant lentivirus-transducing units and 5 mg/ml polybrene (Sigma, USA) were added to each well with gentle mixing to infect the HEK-293T or Tsc2 -/-MEF cells. Cell transfection with miR-24 mimics and inhibitors were conducted using Lipofectamine RNAiMAX (Invitrogen, USA) according to the manufacturer's instructions. For each well, equal doses (100 pmol) of miRNA mimics, inhibitors, siRNAs, or scrambled/negative control RNA were used. All RNA oligoribonucleotides were obtained from GenePharma (Shanghai, China).