Figure 5

Species differences in sex-biased expression of Act79B, an MOL-enriched actin transcript. (a) Quantitative RT-PCR analysis. Genomic DNA (lanes 1 and 2) and first-strand cDNA (lanes 3–6) were prepared from tergites of abdominal segments A3–A6 of males (lanes 1, 3 and 5) and females (lanes 2, 4 and 6) of D. melanogaster, D. subobscura, and 6 species of the montium group (indicated below each panel) for Act79B (lanes 1–4) and 2 control protein genes, α-Tubulin (lane 5) and Act5C (lane 6). M: DL2000 DNA marker. Primers used are as shown in Table S3. (b–d,f–l) in situ hybridization analysis with probes for coding (b–d) or non-coding (f–l) sequences of the Act79B transcript. Act79B expression in abdominal muscles in the wild-type (b,f) and Act79B mutant (d,h) males and wild-type females (c,g) of D. melanogaster and in wild-type males (i,k) and females (j,l) of D. ogumai (i,j) and D. ohnishii (k,l). (e) Phalloidin staining reveals the MOL in a Act79B mutant male of D. melanogaster even though the mutant lacks Act79B expression (d). The true MOLs with Act79B hybridization signals are indicated with arrowheads. The tergite regions typically occupied by the MOL are circled with dotted lines. Oenocytes emit autofluorescence, resulting in a segmentally repeated labelling pattern marked with *. Scale bars: 200 μm (b,e).