Fig. 5. Ectopic expression of RBF delayed proliferation of abdominal histoblasts, but did not result in abnormal numbers of centrosomes during mitosis. White prepupae from either UAS-RBF alone as a control (A) or hsp70-GAL4/UAS-RBF (B-D) were picked and subjected to a 40 minute heat shock at 37°C. Heat shock treatment was repeated every 12 hours until pupae were dissected at 40-43 hours APF. Samples were treated with DAPI to visualize nuclei (A,B, and blue in merged panels), and immunostained with antibodies against ß-tubulin (green in merged panels) and two centrosomal proteins, CP60 and CP190 (red in merged panels). Low magnification views show normal development of the heat shocked UAS-RBF control (A) and the developmental delay caused by ectopic expression of RBF (B). Arrows indicate areas where larval polyploid cells have not been replaced by histoblasts. Representative metaphase (C) and anaphase (D) nuclei from the delayed mitoses have appropriate numbers of centrosomes. Scale bar: in A, 0.1 mm for A,B; in C, 0.01 mm for C,D
