Fig. 4. Suppression of Frizzled-5/7/9 led to female worm ovary morphological alteration.

a RT-qPCR analyses of the transcript levels of Frizzled-5/7/9 in three different regions of the female S. japonicum. Data illustrate representative results showing the mean and standard error, n = 3 independent experiments. Statistical analysis was performed comparing ovary vs anterior or vitellarium using Students t-test and ** denotes P ≤ 0.01. b Inhibition of Frizzled-5/7/9 led to morphological defects of ovaries in females. Female worms were electroporated either with siRNA or control siRNA (3 μg) and then cultured for four days as described in Materials and Methods. S. japonicum worms stained with carmine red and whole-mount preparations of worms imaged by confocal microscopy. Io immature oocyte, mo mature oocyte, v vacuole. The number of parasites similar to representative images is indicated in the upper or lower right of each panel. n = 17–19 parasites, two biological replicates. Scale bar = 20 µm. c Analysis of the ratio of total area of ovary to the area occupied by oocytes as determined by Image J. Data illustrate representative results showing the mean and standard error, n = 6 individual parasites. Statistical analysis compared ovary areas vs areas of appearance of oocytes in ovary using Student’s t test. * P ≤ 0.05 and ** P ≤ 0.01. d Suppression of Frizzled-5/7/9 resulted in decreased egg production. Data illustrate representative results showing the mean and standard error, n = 3 independent wells for egg counts. ** P ≤ 0.01 (student’s t-test, Frizzled-5/7/9 siRNA vs control siRNA).