Figure 6. Bim depletion by short interfering RNA reduces KA-induced neuronal death in hippocampal organotypic cultures.
(A) Representative photomicrographs (4× lens) show control and KA-treated cultures stained with propidium iodide (PI) at 24 h. Note, the significant cell death within hippocampal CA1 and CA3 (arrows). (B) Representative PI staining in KA-treated cultures at 24 h. Note, cell death is reduced in the cultures transfected with short interfering RNA targeting bim (bim siRNA) compared to cultures treated with a scrambled sequence (scram siRNA). (C) Representative PI staining in CA3 for each group. (D) Representative Western blots (n = 3 per lane) showing expression of Bim following KA treatment increases, and that bim siRNA reduces Bim protein levels. Blots below show levels of the KA receptor were not altered by the treatments, and probing for actin is provided as a control for equal protein loading. Data are derived from three independent experiments. (E, F) Graphs show quantification of cell death in CA1 and CA3 24 h after treatment. *p < 0.05 for comparison between cultures treated with KA plus bim siRNA compared to KA-treated cultures treated with scrambled siRNA.