Fig. 6
The BAF complex is required for stem cell access to the epithelial lineage. A ATACseq peaks during RNAi in X1s at loci of key regulators of the epidermal lineage. The sequence coverage tracks show replicate-averaged, sequence depth-normalized (CPM) read coverage for each RNAi condition. B Heatmap of differentially expressed epithelial associated genes during RNAi conditions in X1s. Replicates are displayed independently. C dFISH of piwi-1 and zfp-1 during RNAi at F4D3. Planarians were imaged at the boxed region indicated in the diagram to the left of the images. zfp-1+piwi-1+ cells to total piwi-1+ cells are quantified in graph to the right. Scale bars, 50µm. D Immunostaining of α-PIWI-1 with FISH for nucb1-1 during RNAi. Representative image demonstrating colocalization in control(RNAi) conditions is shown. Planarians were imaged at the boxed region indicated in the diagram to the left of the images and quantified within the nucb1-1 dorsal expression domain. nucb1-1+PIWI-1+ cells to total PIWI -1+ cells in RNAi conditions at F4D3 are quantified in graph to the right Scale bars, 50µm. E Immunostaining of α-PIWI-1 with FISH of p53 during RNAi. Representative image demonstrating colocalization in control(RNAi) conditions is shown. Planarians were imaged at the boxed region indicated in the diagram to the left of the images and quantified within the p53 dorsal expression domain. p53+PIWI-1+ cells to total PIWI-1+ cells in RNAi conditions at F4D3 are quantified in graph to the right. Scale bars, 50µm. F dFISH of late epidermal progenitor marker agat-1 and mature epidermal marker vim-1, as well as Concanavalin-A (ConA), which binds to cell membranes in RNAi conditions, at F4D12. Planarians were imaged in the boxed region indicated in the diagram to the left of the images and dorsal staining was quantified. Representative images are shown (n>8). Scale bars, 50µm. Quantifications are mean \(\pm\) 1 S.D Significance levels in plots: *p<0.05, **p<0.01, ***p<0.001, each “n” is a representative region from 1 planarian