Fig. 2

RNAi knockdown of Smed-bptf phenocopies that of Smed-set1, but not Smed-mll1/2. A) Live images showing the morphological phenotypes of bptf(RNAi) planarians (S.mediterranea) compared to control(RNAi), set1(RNAi), and mll1/2(RNAi) animals. Scale bars = 200 μm. B) Survival plot of set1(RNAi) and bptf(RNAi) animals compared to control RNAi worms. Two non-overlapping constructs are shown for each gene, set1 and bptf, to control for off target effects. C) Whole mount In situ hybridization (WISH) in wild-type worm using a riboprobe to bptf. D) Plots generated from previously published single cell RNA-seq data [7] using the publicly available Shiny App https://simrcompbio.shinyapps.io/bbp_app/ to show bptf expression in specific cell types. E) Schematic of the experimental setup used to test planarian stem cell function in RNAi worms. In normal conditions, a small but significant number of planarian stem cells will survive 1250 rad (12.5 Gy) γ-radiation (3 days post-irradiation, or dpi), then resume proliferating (7 dpi) to restore the population [24, 26, 84]. F) Survival plot of set1(RNAi), mll1/2(RNAi), bptf(RNAi) and control(RNAi) animals with (dotted line) and without (solid line) radiation treatment. G) Representative Fluorescence In situ hybridization (FISH) images of control(RNAi) and bptf(RNAi) planarians stained with the stem cell marker piwi-1. Scale bars = 250 μm. H) Quantitation of piwi-1+ cells per animal for all animals included in E. For each condition (RNAi treatment, +/- radiation, time point) n = 9–12. Statistical significance determined using student’s t-test, *** = p-value < 0.001