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. 2014 May 8;10(5):e1004116. doi: 10.1371/journal.ppat.1004116

Figure 7. SmCBP1 and SmGCN5 play a key role in Smp14 expression and female sexual reproductive development.

Figure 7

A dsRNAi experiment was carried out on sixteen adult worm pairs cultivated for seven days. Worms were electroporated and soaked with dsRNAi from SmCBP1 or SmGCN5 or with the non-specific dsRNAi LUC. On the seventh day of culture, the mRNA levels of SmCBP1, SmGCN5 or Smp14 (graphs A and B) were determined by qRT-PCR, normalized by the α-tubulin transcript levels. The results are depicted in relation to the non-specific dsRNAi (dsLUC) mRNA levels. The effects of the dsRNAi on targeted-protein synthesis were assessed by Western blot (panels C and D). The total number of eggs laid by the same parasites that received the dsRNAi was counted (graphs E and F). Adult worms from the dsRNAi experiments were analyzed by confocal laser scanning microscopy (G). Worms received either the control dsRNAi LUC (panels a and d) or the dsRNAi for SmCBP1 (panels b and e) or SmGCN5 (panels c and f). Panels a–c, and d–f, reveal the effect of the dsRNAi in the ovary, and vitellaria, respectively. OV: ovary; mo: mature oocytes; io: immature oocytes; v: vitellaria; rs: receptaculum seminis. Results are pooled from three independent experiments. Western blots were repeated three times. The confocal microscopy images are representative of several parasites analyzed. Scale bars 10 µm. Student's t-test was applied, with *p<0.05, **p<0.01, ***p<0.001 and ****p<0.0001.