Fig. 3: Ectopic expression of the grh.RP (N-class isoform) promotes differentiation.
From: Alternate Grainy head isoforms regulate Drosophila midgut intestinal stem cell differentiation
Representative images of control midguts (A), midguts overexpressing Grh.RP (B), the apoptotic inhibitor P35 (C) and midguts co-expressing P35 and Grh.RP (D) in progenitor cells. Scale Bar 40 μm. E Quantification of the number of progenitor cells in control (n = 15 midguts) midguts, midguts over expressing Grh.RP (n = 19 midguts), P35 (n = 22 midguts) and midguts co-expressing Grh.RP and P35 (n = 7 midguts) shows that expression of p35 was not able to rescue loss of progenitor cells. (Mean ± SEM, One-Way ANOVA with Tukey’s Test, ns not significant, ****p < 0.0001). F Quantification of the proportion of ISC/EBs in control and esgTS > UAS-grh.RH (N′ isoform) midguts. The proportion of progenitors in esgTS > UAS-Grh.RH midguts (n = 21) have decreased in comparison to control midguts (n = 13). (Mean ± SEM, Unpaired Student's T-test, **p = 0.0088). Confocal images of control (G) and midguts ectopically expressing Grh.RP (H) and Grh.RH (I) and Grh.RL (Grh.O-class) (J) in ISC/EBs. Compared to controls, GFP+ progenitor cells expressing Grh.RP, Grh.RH or Grh.RL have increased cell size and also express the EC marker, Pdm-1 (arrows). Scale Bar 40 μm. K The proportion of esg+ progenitor cells expressing Pdm-1 in midguts increases in midguts over expressing N-class isoforms Grh.RP (n = 15), Grh.RH (n = 9) and O-class isoform Grh.RL (n = 8) in comparison to control midguts (n = 11). While the proportion of esg+ progenitor cells expressing Pdm-1 slightly increased in midguts overexpressing Grh.RH (n = 19), it was not statistically significant compared to controls. (Mean ± SEM, one-way ANOVA with Tukey’s Test, ns not significant, *p = 0.0107, ****p < 0.0001).
