Figure 5.

The silencing of BIK decreased cell death induced by Len/VD3. (a–d) MCL cells (5 × 10⁵/ml) were seeded for 48 h with or without Len/VD3 prior or not to transfection with siControl (siCt) or siBIK RNA, and the cells were reseeded (5 × 10⁵ cells/ml) for an additional 3 days with or without Len/VD3. (a) Western blotting analysis of Bik expression. Bik expression was assessed in siCt and siBIK MCL cells that were or were not treated with combined Len/VD3. A representative experiment out of three is shown. (b) The silencing of BIK prevented the Len/VD3-induced cell growth inhibition. The cellular density was determined by direct counting. The data represent four independent experiments. (c) The silencing of BIK decreased Len/VD3-induced death. The cells were stained with Annexin V. The data represent four independent experiments. (d) The silencing of BIK decreased Len/VD3-induced subG1 peak. The cells were stained with PI. The data represent four independent experiments. (e–g): JEKO-1 cells (5 × 10⁵/ml) were seeded for 48 h with or without Len/VD3 prior or not to transfection with siCt or siNOXA or siPUMA RNA, and the cells were reseeded (5 × 10⁵ cells/ml) for an additional 3 days with or without Len/VD3. The data represent three independent experiments. (e) Western blotting analysis of Puma and Noxa expression in JEKO-1 cells. A representative experiment out of three is shown. (f) SiNOXA and siPUMA RNA did not prevent Len/VD3-induced inhibition of growth. The cellular density was determined by direct counting. The data represent three independent experiments. (g) SiNOXA and siPUMA RNA did not inhibit Len/VD3-induced cell death. Cells were stained with Annexin V. The data represent three independent experiments. *P<0.05, **P<0.01. NS, not significant