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. 2022 Dec 12;45(12):886–895. doi: 10.14348/molcells.2022.2031

Fig. 3. RUNX1 directly transactivates survivin expression.

Fig. 3

(A) Proximal regulatory region (−1,500 to 0 bp relative to the transcription start site [TSS]) of Survivin. (B) Result of the chromatin immunoprecipitation (ChIP) analysis of KP-MRT-YM cells using anti-RUNX1, an isotype-matched control IgG, and anti-Histone H3 antibodies. ChIP products were amplified via polymerase chain reaction (PCR) to determine the abundance of the indicated amplicons. (C) Luciferase reporter assay with survivin promoter. HEK293T cells were stably-transduced with the lentivirus expressing RUNX1 (RUNX1 o/e) or control, together with the reporter vector expressing luciferase gene under survivin promoter. Cells were incubated with 3 μM doxycycline for 48 h, and the luciferase activity was monitored with a luminometer. Result was normalized to that of the control sample (n = 3). (D) Immunoblotting of RUNX1, survivin, and GAPDH in non-depleted and RUNX1-depleted (RUNX1 K/D) MP-MRT-AN cells transduced with or without lentivirus expressing survivin (Survivin o/e). Cells were treated with 3 μM doxycycline for 48 h and lysed for immunoblotting. (E) Restoring survivin expression in RUNX1-depleted MP-ART-AN cells reverts RUNX1-depletion-mediated growth inhibition. The indicated cells were cultured in the presence of 3 μM doxycycline (n = 3). Data are represented as the mean ± SEM. *P < 0.05, by two-tailed Student’s t-test.