Fig. 5

Mosaic expression of egfp regulated by 5 × UAS and methylation analysis of 5 × UAS-E1b by bisulfite sequencing.

(a) Schematic of the expression gene cassette of the plasmids used to establish the TG6 line. (b) Variable mosaic expression of the 5 × UAS-regulated egfp gene was detected in the offspring of the TG4 and TG6 lines (at 60 hpf). The variable mosaic expression of the 5 × UAS-regulated egfp gene was detected in (c) the testes and (d) the ovaries of the offspring of the TG4 and the TG6 lines (at 3 months of age). (Scale bars: 100 µm). The expression of dnd in (e) the male gonads and (f) female gonads of the WT, TG3-1 and TG3-2 lines was validated by quantitative PCR (Q-PCR). The expression of gal4 in (g) the male gonads and (h) the female gonads of TG3-1 and TG3-2 individuals was validated by Q-PCR. The 2^-?Ct values were plotted as the relative mRNA level normalized to odc1. Values are represented as the mean ± SEM of four repeats. *P < 0.05, ** P < 0.01. (i) DNA from the gonads of the TG3-1 and TG3-2 adults (at 4.5 months of age) was subjected to bisulfite sequencing. CpG methylation patterns in the 5 × UAS-E1b are indicated on the horizontal axis. The first ten CpG sites resided within the 5 × UAS, the last two CpGs sites resided within the E1b promoter. Ten clones were tested and the results are shown on the vertical axis. Black circles represent methylated sites.