FIG. 3.

Bim expression involved in luminal apoptosis and luminal clearance during morphogenesis. (A) At 24 h after transfection with Bim siRNA oligonucleotides or matched control oligonucleotides, cells were placed in morphogenesis assays, and cell lysates were collected at the indicated times. Protein levels were analyzed by immunoblotting with Bim or actin antibodies. (B) Cells transfected with Bim or matched control siRNA oligonucleotides were placed in a morphogenesis assay, and the percentage of acini with EtBr-positive cells (two or more cells) was scored after the indicated number of days in culture. Values represent the mean ± the SEM of four independent experiments. (C) Cells transfected with Bim or matched control siRNA oligonucleotides were placed in morphogenesis and were fixed and immunostained at day 10 with anti-active caspase-3 (green), and nuclei were counterstained with TOPRO-3 (blue). Scale bar, 20 μm. (D) Nuclei of representative day 12 structures from cells transfected with Bim or matched control siRNA oligonucleotides were stained with TOPRO-3 (red). Scale bar, 20 μm.