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. 2001 Oct 15;15(20):2741–2754. doi: 10.1101/gad.916201

Figure 5.

Figure 5

Over-expression of Dm Geminin during Drosophila development reduces DNA replication. (A) A stage-11 wild-type embryo with BrdU (red) and anti-En (green in A–L). (B) A stage-11 en–GAL4 UAS–Dm geminin embryo with BrdU (red) and anti-En showing decreased BrdU labeling in the En stripes. (C) A wild-type stage-11 embryo stained with anti-En and PI (red). (D) A stage-11 en–GAL4 UAS–Dm geminin embryo stained with anti-En and PI (red). (E) High magnification of A. (F) High magnification of B. Arrows indicate En stripes. (G) High magnification of C. (H) High magnification of D. (I,J) High magnification of stage-11 wild-type embryo with anti-En and anti-PH3 (red). (K,L) High magnification of stage-11 en–GAL4 UAS–Dm geminin embryos with anti-En and anti-PH3 showing more PH3 in En stripes. (I–L) Anaphase (a); prophase cells (p); and metaphase cells (m) within the En stripe are indicated. An average of 14.1±4.1 (SE) mitotic cells were present in the En stripe in the en–GAL4 UAS–Dm geminin embryo compared with an average of 3.5 ± 1.2 (SE) in an equivalent sized interstripe region (n = 13). Wild-type embryos contained a similar density of mitotic cells in En stripes compared with interstripe regions. (M) High magnification of TUNEL (blue) and En (red) in a wild-type stage-11 embryo. (N) High magnification of TUNEL (blue) and En (red) in an en–GAL4 UAS–Dm geminin stage-11 embryo showing more TUNEL-positive cells. (O–Q) BrdU labeling of eye imaginal discs from wild type (O), GMR–GAL4 UAS–Dm geminin (3 copies) (P), and ey–GAL4 UAS–Dm geminin (2 copies) (Q). In P and Q the brackets indicate the regions where Dm geminin is ectopically expressed. Wild-type eye discs contained an average of 119±14 (SE) (n = 3) compared with 63±6 (n = 4) S-phase cells for GMR–GAL4 UAS–Dm geminin eye discs within an equivalent region of the post-MF S-phase band. The MF is indicated (*). (R–V) Scanning electron micrograph (SEM) of adult eyes from (R) ey–GAL4 UAS–Dm geminin (2 copies), (S) wild type, (T) GMR–GAL4 UAS-Dm geminin (3 copies), (U) GMR–GAL4 UAS–Dm geminin (2 copies), (V) GMR–GAL4 UAS–Dm geminin (2 copies)/dupa1. Note in V that the eye is smaller and more disorganized than in U. Adult eyes are oriented anterior to the right.