Fig. 2: grhWG MARCM clones and O-class isoform-specific RNAi also result in loss of progenitor cells.
From: Alternate Grainy head isoforms regulate Drosophila midgut intestinal stem cell differentiation
Confocal images of control (A) and grhWG (B) MARCM clones maintained at 25 C. Scale Bar 40μm. C, C′ Quantification of the number of clones per midgut in control (6DPCI: n = 18 midguts 10DPCI: n = 26 midguts) and grhWG (6DPCI: n = 15 midguts, 10DPCI: n = 26 midguts) MARCM clones at 6DPCI and 10DPCI. A reduction in the number of clones in grhWG midguts is observed at both time points quantified. (Mean ± SEM, Unpaired Student's T-test, ****p < 0.0001). D, D′ Quantification of clonal size between control and grhWG MARCM clones. At both 6DPCI and 10DPCI, grhWG clonal size, measured by counting the number of cells per clone is smaller than that of control clones. Control – 6DPCI: n = 754 clones, 18 midguts; 10DPCI: n = 1169 clones, 26 midguts. grhWG – 6DPCI: n = 242 clones, 15 midguts; 10DPCI: n = 499 clones, 26 midguts. (Mean ± SEM, Unpaired Student's T-test, ***p = 0.0003, ****p < 0.0001). E, F′ Confocal images of control and esgTS > UAS-grh.O-class RNAi midguts. ISCs are marked by Dl and progenitor cells marked with GFP. Scale Bar 40 µm. Inset images are magnified views of regions enclosed by dotted boxes. G Quantification of Dl+ ISC cells in control (n = 22 midguts) and esgTS > UAS-grh.O-class RNAi (n = 24 midguts) midguts showed a decrease in the proportion of Dl+ cells over total cell number. (Mean ± SEM, Unpaired Student's T-test with Welsh’s Correction, **p = 0.0078). H Proportion of esg+ cells that are Dl+ ISCs in control (n = 22 midguts) and esgTS > UAS-grh.O-class RNAi (n = 24 midguts). (Mean ± SEM, Unpaired Student's T-test with Welsh’s Correction, **p = 0.0049). I Quantification of the proportion of Dl+ ISCs over total cell number in control (n = 12 midguts) and esgTS > UAS-grh.ALL RNAi (n = 18 midguts) show no significant difference between the two genotypes. (Mean ± SEM, Unpaired Student's T-test with Welsh’s Correction, ns not significant). J The proportion of esg+ cells that are Dl+ ISCs remain at similar levels in control (n = 12 midguts) and esgTS > UAS-grh.ALL RNAi (n = 18 midguts) midguts. (Mean ± SEM, Unpaired Student's T-test with Welsh’s Correction, ns = not significant).
