Figure 7
From: HDAC inhibitors induce tumor-cell-selective pro-apoptotic transcriptional responses
Forced expression of BFL-1 protects BJ LTSTERas fibroblasts from vorinostat-mediated apoptosis. (a) Proteins were extracted from exponentially growing BJ LTSTERas fibroblasts, and MSCV and MSCV FLAG–BFL-1 transduced variants. Proteins were separated on 10% polyacrylamide gels by SDS-PAGE and analyzed for the expression of FLAG–BFL-1 by western blotting with an anti-FLAG antibody. The expression of α-Tubulin levels served as a control for protein loading. (b) BJ, BJ LTSTERas, BJ LTSTERas MSCV and BJ LTSTERas FLAG–BFL-1 cells were incubated with 0, 2.5, 5, 10, 25 and 50 μM vorinostat for 24, 48 and 72 h. Cells were harvested and stained with annexin-V-APC and propidium iodide as an apoptosis readout or stained for loss of mitochondrial outer membrane potential (MOMP, ΔΨm) using the mitochondrial dye tetramethylrhodamine ethyl ester (TMRE). Cells were then analyzed by flow cytometry. The percentage of annexin-V-APC, propidium iodide double positive cells are shown in the left panels while cells that had lost the ability to bind TMRE (ΔΨm) are in the right panels. Data are presented as mean±S.E.M. of three independent experiments
