Fig. 3. Overexpression of Crol promotes cell cycle. (A,B) BrdU (red) of control en-GAL4, UAS-GFP/+. (C,D) BrdU of en-GAL4, UAS-GFP/UAS-Crol. (E,F) PH3 (purple) of control. (G,H) PH3 of the same disc for which BrdU is shown in C and D. In A-H, the arrow indicates the ZNC. (I,J) TUNEL labelling (red) of the basal layer of an en-GAL4, UAS-GFP/UAS-Crol disc and counterstained with DAPI (blue) in I. (K) en-GAL4, UAS-GFP/+;UAS-p35/+ control and (L) en-GAL4, UAS-GFP/UAS-Crol;UAS-p35/+. Discs in K and L are both taken at 10x magnification and stained with PH3 (red) and DAPI (blue). (M-P) 120-hour en-GAL4, UAS-GFP/UAS-Crol;UAS-p35/+ taken at 20x magnification compared with 40x for discs in A-H. (M) BrdU (red), (O) PH3 (purple). The merge with GFP and DNA (blue) is shown with BrdU in N and PH3 in P. In M-P, a white line marks the AC of the pouch. (Q-X) Cell cycle analysis of 120-hour discs with flip-out clones 72 hours after induction. (Q) UAS-p35 with BrdU (red), (R) UAS-crol;UAS-p35 with BrdU (red) and merged with DAPI in S. (T) Quantification of BrdU in 120-hour discs. Counts are from equivalent GFP-positive areas (three sets of 70,000 pixels) for UAS-p35 (249.7±14.2) and UAS-crol;UAS-p35 (509.8±10.9). (U) UAS-p35 with PH3 (purple), (V) UAS-crol;UAS-p35 with PH3 (purple) and merged with DAPI in W. (X) Quantification of PH3 (as above) from UAS-p35 (39.9±0.5) and UAS-crol;UAS-p35 (67.9±7.7). A statistically significant increase was observed between UAS-crol;UAS-p35 and UAS-p35 clones for BrdU (P<0.0001) and PH3 (P<0.0033).