Figure 7: Schistosome egg development and ovarian architecture are regulated by cytosine methylation.

(a) 5-Azacytidine (5-AzaC) inhibits egg production in a dose-dependent manner. Histograms represent mean (+s.e.m.) egg output derived from five worm pairs (n=4) cultured for 48 h. A one-way analysis of variance (ANOVA) followed by post hoc testing with Fisher's Least Significant Difference (LSD) was used to detect statistical differences (*P<0.05) in egg production among groups (control, 10 μM 5-AzaC and 100 μM 5-AzaC). Data are derived from a representative experiment (replicated five times). (b) Confocal microscopy of schistosome eggs laid by control worm pairs (control) or 5-AzaC-treated worm pairs (5-AzaC, 100 μM). Scale bar, 20 μm. (c) Confocal microscopy of normal adult females (control) or 5-AzaC-treated females (5-AzaC, 100 μM) stained with Langeron's Carmine and imaging focused on a region containing the intra-uterine egg. Scale bar, 20 μm. (d) 5-AzaC inhibits egg development as assessed by in vitro staging. Eggs deposited by control females (control) or 5-AzaC-treated females (5-AzaC, 100 μM) were left to develop as described in the Methods section and maturation classified according to four categories (stage I+II, stage III−V, hatched or unidentified; scale bar, 20 μm). The percentage of eggs displaying each developmental stage is indicated by a stacked histogram and is representative of two independent experiments. (e) Confocal microscopy of normal adult females (control) or 5-AzaC-treated females (5-AzaC, 100 μM) stained with Langeron's Carmine and imaging focused on a region containing the ovary. Scale bar, 20 μm. Io, immature oocyte; mo, mature oocyte; v, vacuole.