Fig. 5: Wnt signalling functions in the Smed reproductive system.
From: Evolutionary dynamics of whole-body regeneration across planarian flatworms
a, Co-occurrence of robust head regeneration (Group A, red) with fissiparous reproduction across planarian clades. b, Live images illustrating the sexual (egg-laying; left) and asexual (fissiparous; right) reproduction modes of Smed laboratory strains. c, Cartoon (top) and colorimetric whole-mount in situ hybridizations (indicated markers; middle) of Smed sexual strain reproductive system components; reproductive system ablation under ophis(RNAi) (bottom). d, ß-CATENIN-1 amounts/Wnt signalling activity in A–P sections (1, head; 6, tail) quantified via Western blotting with the G78 mAB. Smed strain and RNAi conditions as indicated. Error bars, s.d. of n = 4 biological replicates, each representing the mean of four technical replicates (blots). e, Wnt component expression in the Smed reproductive system by colorimetric or fluorescent whole-mount in situ hybridization. Number pairs, specimen fraction displaying the pattern shown; red arrowhead, testes lobules; black arrowhead, expression in non-reproductive tissues; H3P, Histone H3 Ser10 phosphorylation immunolabelling; DAPI, nuclei. f, Colorimetric whole-mount in situ hybridizations of the indicated reproductive system markers under the indicated RNAi conditions; number pairs, specimen fraction displaying the pattern shown; dashed lines, approximate position of the sagittal sections in Fig. 5g. g, Relative yolk gland cross-sectional area quantifications in histological sections of the indicated RNAi conditions. Top, bar graph, n = 2 individuals (dots)/condition; error bars, s.d. of the mean. Bottom, representative Mallory-stained sagittal sections. White outline, yolk glands; red boxes, zoom views. y, yolk; vnc, ventral nerve cord. h, Coomassie-stained SDS–PAGE of lysates of the indicated sources. White box, major yolk protein. i, Fluorescent Western blot of the indicated lysates, probed with the indicated antibodies. j, EO95 specificity analysis in Smed sexual strain lysates under the indicated ferritin(RNAi). Asterisk, EO95 signal loss in ferritin-C(RNAi). k, Control (eGFP)-normalized quantification of FERRITIN-C and ß-CATENIN-1 amounts in lysates of the indicated RNAi condition via EO95 and G78 immunoblotting. Error bars, s.d. of the mean of four technical replicates (dots) in n = 1 biological replicate. l, Colorimetric whole-mount in situ hybridization of the four yolk ferritins in sexual Smed under the indicated RNAi treatments. Scale bar, 1 mm unless otherwise noted.
