Fig. 2: Separase regulates nuclear lamin protein levels.

Western blot analysis of total protein extracts from wild-type Oregon-R (Or-R) and Sse^dft mutant (dft) third instar larval neuroblasts, probed with anti-Dm0 (A) and anti-LamC (B) antibodies. Anti-Actin antibody was used as a loading control. Quantification of Dm0 (C) and LamC (D) protein signals from western blot analysis. Data were collected from three independent experiments (*p < 0.05, **p < 0.01; Student’s t test). qPCR quantification of Dm0 (E) and LamC (F) mRNA expression. At least three independent experiments were conducted (*p < 0.05, **p < 0.01; Student’s t test). Western blot analysis of FLAG-tagged LamC expression in wild-type (Tg LamC) and Sse^dft mutant (Tg LamC dft) backgrounds (G), with quantification of transgenic LamC expression levels (H). Three independent experiments were performed (*p < 0.05, **p < 0.01; Student’s t test). anti-Giotto (GIO) was used as a loading control. Immunofluorescence staining with anti-Dm0 (I) and anti-LamC (J) antibodies (green) in intact nuclei from Or-R and Sse^dft third instar larval salivary glands. DAPI was used to stain DNA. Note the presence of LamC aggregates in the Sse^dft nuclei (arrows). Quantification of Dm0 (K) and LamC (L) fluorescence intensity from salivary gland immunostaining (***p < 0.001; Student’s t test). M Immunostaining with anti-cleaved caspase Dcp-1 (red) in larval brains from untreated and IR-treated (10 Gy) Oregon-R controls, and from untreated (dft) and IR-treated (10 Gy dft) Sse^dft mutant brains.