Fig. 9: In situ HCR expression of proliferation and differentiated cell markers in piwi+ cells.

A In situ HCR expression of piwi+ cells marker PrileiEVm022498t1 (histone h3, magenta) and EdU+ cells (yellow) showing signal throughout the whole animal body. The right microscopy panels are close-ups from different animals, showing overlapping expression in the trunk, fission zone (FZ), and posterior growth zone (PGZ). The bottom right microscopy panel is a close-up from the upper right microscopy panel, evidencing the overlapping expression in a cell in the FZ. All cells were stained with DAPI (grey). B In situ HCR and expression plot of piwi+ markers PrileiEVm022498t1 (histone h3, magenta) and PrileiEVm003567t1 (piwi1, green), and EdU+ cells (yellow), showing extensive signal in the PGZ. The middle and bottom microscopy panels are close-ups from the upper microscopy panel evidencing overlapping expression in the PGZ and at the cellular level. Dashed line indicates the outline of the worm. C In situ HCR and expression plot of piwi+ marker PrileiEVm022498t1 (histone h3, magenta) and gut marker PrileiEVm022781t1 (cyan), and EdU+ cells (yellow), showing expression in the developing gut of the new worm that has just split apart. The middle and bottom microscopy panels are close-ups from the upper microscopy panel evidencing overlapping expression in the gut and at the cellular level. D In situ HCR and expression plot of piwi+ marker PrileiEVm022498t1 (histone h3, magenta) and neural and polycystin marker PrileiEVm025662t1 (cyan), and EdU+ cells (yellow), showing intensive expression in the developing brain in FZ. The middle and bottom microscopy panels are close-ups from the upper microscopy panel evidencing overlapping expression in the developing brain and at the cellular level. E In situ HCR and expression plot of piwi+ marker PrileiEVm022498t1 (histone h3, magenta) and epidermal marker PrileiEVm008287t1 (intermediate filament, cyan), and EdU+ cells (yellow), showing intensive expression in the FZ. The middle and bottom microscopy panels are close-ups from the upper microscopy panel evidencing overlapping expression in the epidermis and at the cellular level. In all panels, anterior is left, dorsal is up. Scale bars are 50 μm unless noted in the figure. All expression patterns displayed in the figure were observed in, at least, three different individuals.