iSLK/RTA-WT and iSLK/RTA-DD stable cell lines were established using Tet-On 3G Tetracycline Inducible Gene Expression Systems. Briefly, 293T cells were transfected with the packaging plasmids VSV-G, Gag-pro, Vpr-pol, Tet-off, and tat-IRES-rev and the TetONE-RTA-WT or TetONE-RTA-DD lentiviral expression vector. For making shRNA lentivirus, 293T cells were cotransfected with packaging plasmids VSV-G, DR8.9, and the shRNA lentiviral plasmid as previously described (53). At 48 hours after transfection, the supernatant was harvested and filtered, and viruses were concentrated by ultracentrifugation. SLK or 293T cells were spin-infected (1800 rpm) with lentivirus in the presence of polybrene (8 μg/ml) for 45 min. Cells were selected with puromycin (1 μg/ml) at 48 hours after infection.

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