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. 2015 Dec 1;9(12):e0004258. doi: 10.1371/journal.pntd.0004258

Fig 4. Suppression of expression of CYP in adult Opisthorchis felineus by RNA interference (RNAi).

Fig 4

A. Transcript levels were determined using EVA-green real-time RT-PCR. The MrpL16 gene was used for normalization. The control group (no dsRNA treatment) was compared to the negative control group transformed with LUC dsRNA. Three biological samples with technical duplicates were used for analysis. The data are presented as means ± SD. B. Primer positions for analysis of CYP gene expression. C. Control adult O. felineus, eight days after electroporation. EC: excretory channel, EB: excretory bladder. D. Worms eight days after of the knockdown of CYP mRNA. E. Percentage of worms with phenotypic changes in excretory system after three days of RNA interference (data of three independent experiments). Wild type–untreated worms; mock–worms subjected to electroporation without dsRNA; LUC–worms that received LUC dsRNA (non specific control); CYP–worms that received CYP dsRNA; keto—worms were treated with ketoconazole for three days. F, G. Worms five days after the knockdown of CYP (F) and LUC (G) mRNA were treated for 20 h with pentoxyresorufin, and images acquired using multiple fluorescence filters using optical sectioning by means of the AxioImager fluorescence microscope (Zeiss). The resorufin (R) forming in the excretory bladder is indicated with an arrow. Representative images are shown. H. Adult O. felineus after three days of treatment with 40 μM ketoconazole.