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Fig. 1. SOP is arrested at G2 phase in phyl mutants. (A-K) SOPs were labeled by anti-Sens staining (green). (A,B) Wild-type pupal thoraces at 14 hours (A) and 16 hours (B) APF. Almost all wild-type SOPs have divided within these 2 hours. (C,D) SOP division is delayed in phyl² mutant clones (GFP negative). Arrows indicate phyl²-mutant SOPs, which remain as single cells at 22-24 hours APF (C) and do not divide until 24-28 hours APF (D). (E,G,I) Wild-type pupal thoraces. (F,H,K) phyl² mutant clones. (J) phyl²/phyl⁴ hypomorphic mutant. (E-F') Thoraces stained with anti-PH3 antibody in red. Yellow and white arrows indicate the wild-type mitotic and phyl² mutant SOPs, respectively. (G-H') Thoraces labeled for CycB expression in red. Yellow and white arrows indicate the wild-type and mutant SOPs, respectively. All SOPs accumulate high levels of CycB. (I-J') Thoraces labeled for BrdU incorporation in red. BrdU signals are observed in the two sister cells resulting from the first division of the SOP (indicated by arrows), but are never detected in wild-type SOPs (I) and in phyl²/phyl⁴ hypomorphic mutant SOPs (J). (K,K') BrdU incorporation assay in phyl² null mutant clones. SOP and its lineage cells were labeled by anti-Sens (green), and BrdU signals are shown in red. The broken lines mark the boundary of phyl² mutant clone (GFP negative). Although both anti-Sens staining and GFP signals are shown in green, anti-Sens staining signals are stronger than GFP and are arranged in regular arrays. The uniform GFP signals are weaker due to HCl hydrolysis used in the BrdU labeling procedures (see Materials and methods). BrdU signals can be easily detected in neighboring wild-type SOP-lineage cells (indicated by yellow arrows), but not in phyl² mutant SOPs (indicated by white arrows).

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