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Fig. 2. crol is required for cell cycling. (A-N,P-S) Clones
generated using MARCM (positively marked with GFP). (A,B)
120-hour wild-type clones; (C,D) 120-hour
crol-/- clones. (E) 120-hour crol mutant
clones expressing UAS-crol. (F-J) crol clones die by
apoptosis. (F) Apical section of 96-hour crol clones.
(G) Basal section of the disc in H, counterstained with propidium
iodide (red). (H) Higher magnification to show pyknotic nuclei with
DAPI (blue) and (I) merged with GFP. (J) 120-hour
crol-/- clones overexpressing UAS-p35. In B and
D, discs have been co-stained with phalloidin-TRITC to distinguish the hinge
from the pouch; the pouch is marked by a white line in A,C,E-G,J. (K-T) Cell
cycle analysis of crol mutant clones in the UAS-p35
background. (K-N) 96-hour discs 48 hours after heat-shock. BrdU
labelling (red) in control clones (K) and crol-/- clones
(L) or PH3 staining (purple) in control (M) and crol mutant clones
(N). (P,Q) crol-/- clones in 120-hour discs
with BrdU (P) and a higher magnification (Q) to show cells flanking the ZNC,
which corresponds to the white square in P. (R,S)
crol-/- clones in 120-hour discs with PH3 and a higher
magnification (S) to show mitotic cells adjacent to the ZNC, which corresponds
to the white square in R. (O,T) Quantification of BrdU (O) and
PH3 (T) in 96-hour discs. Counts are from equivalent clone areas (three sets
of 70,000 pixels) from control (BrdU 488.6±24.3, PH3 52.0±3.2),
crol-/- (BrdU 84.24±5.2, PH3 7.02±0.6) and
crol-/-; UAS-p35 (BrdU 131.61±7.8, PH3
22.14±2.4). A statistically significant decrease was observed between
crol-/- and control clones for BrdU (P<0.0001)
and PH3 (P<0.0001), and for crol-/-;
UAS-p35 and control clones BrdU (P<0.0001) and PH3
(P<0.0002).
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