Fig. 3: An RNAi screen identifies that boule phenocopies oc-1 after RNAi in the juvenile male germline.

a WISH images showing the expression of boule, akkn, lsm14, zc3h31, ccnb1ip1, and uaf in male juvenile parasites. Asterisks: testes. The imaged areas correspond to the dashed box in the schematic on the left. N: number of samples showing similar results over two independent experiments. b Confocal images showing representative individual testis lobules stained by DAPI and EdU in control and boule, akkn, lsm14, zc3h31, ccnb1ip1, and uaf RNAi juvenile parasites. Insets: magnified boxed areas. Dashed circles: testis lobule boundary. Arrowheads in ccnb1ip1 RNAi image: incomplete separation of meiotic nuclei. Nuclear morphologies are labeled in the images as in Fig. 2c. c Fraction of EdU⁺ nuclei, d number density of GSCs and e fraction of differentiated germ cells in testes in control and RNAi parasites. Each data point represents the mean across all testis lobules in a single parasite. N = 12 (control RNAi); N = 12 (oc-1 RNAi), N = 12 (boule RNAi), N = 4 (akkn RNAi); N = 13 (lsm1 RNAi); N = 6 (zc3h31 RNAi); N = 6 (ccnb1ip1 RNAi). Data are plotted as average ± standard deviation and p-values are calculated using two-sided Welch’s t-test. f, g FISH images showing broader expression of nanos-1 (f) and eled (g) in testes after boule RNAi. Arrowheads: differentiated germ cells that do not express nanos-1 or eled. n: number of samples exhibiting the reported phenotype out of the total number of samples analyzed. RNAi experiments were repeated on at least three biological replicates.