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. 2004 Oct 14;101(43):15313–15317. doi: 10.1073/pnas.0406837101

Fig. 1.

Fig. 1.

The expression and distribution of BIM are under the control of IL-3. (A) Small interfering RNA knockdown of BIM expression protects from apoptosis induced by IL-3 deprivation. (Upper) IL-3 was withdrawn from FL5.12 cells transfected with either pSuper-BIM (small interfering BIM) or pSuper (control). Total cell extracts were subjected to Western blot with anti-BIM. (Lower) Cell viabilities were determined at the indicated time points by Annexin V-PI staining. Values represent the mean ± SD of three experiments. (B) IL-3 was withdrawn from FL5.12 cells for the indicated times. Total cell extracts were subjected to Western blots with anti-BIM. Arrows indicate the position of BIM isoforms, BIMEL and BIML, respectively (Upper). IL-3 was withdrawn from FL5.12 cells for 4 h, followed by readdition of IL-3 for the indicated times (Lower). (C) FL5.12 cells cultured in IL-3 (time 0), IL-3 deprived for 8 h (-IL-3), and deprived cells reexposed to IL-3 for 15 min (+IL-3) are shown. Cells were fractionated into cytosol (Cyt), heavy membrane (HM), and light membrane (LM) fractions. Western blots were performed with anti-BIM, anti-BAX, and anti-VDAC1.