Fig. 6: Pax regulates midgut homeostasis dependently on Yki-Sd.
a–d’ Adult midguts of Esgts>Ctrl (a–a’), Esgts>Pax RNAi-1 (b–b’), Esgts>Pax RNAi−1;Yki RNAi-1 (c–c’) and Esgts>Pax RNAi-1;Sd RNAi (d–d’) were immunostained with p-H3+(red) and DAPI (nuclei, blue). ISCs and EBs/pre-EEs are marked with Esg-GFP (green). White arrows indicate the p-H3 signal. e Quantification of p-H3+ cells of adult midguts of the indicated genotypes of (a–d’) and (Supplementary Fig. 11a–f’) (n = 10,13,15,10,11,10,10,10,10). The counting of p-H3+ cells was conducted across the entire midgut. f–i’ Adult midguts containing GFP-positive MARCM clones of Ctrl (f–f’), sdΔB1 (g–g’), Pax RNAi (h–h’), sdΔB1 in the presence of Pax RNAi (i–i’) were dissected 3 days after clone induction and immunostained with Arm+Pros (red) and DAPI (nuclei, blue). Dotted lines mark the edges of the clone region. j Quantification of the cell number per clone in adult midguts from the indicated genotypes in (f–i’) at 3 days after clone induction (n = 30, 70, 33, 35). k–n’ Adult midguts containing GFP-positive MARCM clones of Ctrl (k–k’), ykiB5 (l–l’), Pax RNAi (m–m’), ykiB5 in the presence of Pax RNAi (n–n’) were dissected 3 days after clone induction and immunostained with Arm + Pros (red) and DAPI (nuclei, blue). Dotted lines mark the edges of the clone region. o Quantification of the cell numbers per clone in adult midguts from the indicated genotypes in (k–n’) at 3 days after clone induction (n = 35, 40, 46, 80). p–s’ Relative wing discs containing GFP-positive Flip-out clones of Ctrl (p–p’) and Pax RNAi (q–q’), Yki RNAi (r–r’), and Pax RNAi;Yki RNAi (s–s’) were dissected 3 days after clone induction and immunostained with DAPI (nuclei, blue). t Quantification of cell number of clones in panels (p–s’) (n = 14 for each genotype). Three independent experiments were performed, and the error bars are mean ± SEM. In each box plot, the center line indicates the median, the edges of the box represent the first and third quartiles, and the whiskers extend to the minimum and maximum values. P values of significance (indicated with asterisks, NS no significance P ≥ 0.05, *P < 0.05, **P < 0.01, ***P < 0.001, ****p < 0.0001) were calculated by one-way ANOVA with Tukey’s test (e, j, o, t). Scale bars: 30 µm (a–d’, f–i’, k–n’), 60 µm (p–s’). Confocal images were taken from the boundary region between R4c and R5a of the posterior midgut.
