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. 2012 Jun 15;7(6):e39117. doi: 10.1371/journal.pone.0039117

Figure 3. siRNA targeting BFL-1 but not MCL-1 diminishes activation-induced survival of human mast cells.

Figure 3

(A) The upregulation of BFL-1 and MCL-1 following IgECL in CBMCs is abolished following targeted siRNA treatment as verified 30 hours post-transfection by quantative PCR. Cells were transfected with 100 nM siRNA, sensitized with 1 μg/mL of IgE and challenged with 2 μg/mL anti-IgE before expression was determined. Data correspond to one representative experiment using one donor. Similar result was obtained for another donor. (B) BFL-1 but not MCL-1 siRNA treated CBMCs show decreased survival upon IgECL compared to control siRNA treated cells. Cells were transfected, sensitized with 1 μg/mL of IgE and 24 hours post-transfection cytokine-deprived and challenged with 2 μg/mL anti-IgE before being enumerated 24 hours later using the vital dye trypan blue. N=6. (C) The upregulation of BFL-1 and MCL-1 following IgECL in LAD-2 cells is abolished following targeted siRNA treatment as described above. (D) BFL-1 but not MCL-1 siRNA treated LAD-2 cells show decreased survival upon IgECL compared to control siRNA treated cells. N=4. Change in viability is percentage viable cytokine-deprived cells deducted from viable cytokine-deprived cells following IgECL. (E) Bfl-1 is down-regulated in LAD-2 cells following siRNA treatment targeting BFL-1 as compared to control siRNA in immunohistochemical stainings for Bfl-1 expression.