Supplementary Figure 1: Reporter constructs and apicobasal polarity in Ngn3 cells at E15.5 and E12.5.
(a) Construct of the Ngn3RG1 reporter. (b) Representative section of an E15.5 Ngn3RG1 pancreas, stained for Neurogenin3 (Ngn3; white) and E-cadherin (E-cad; blue) shows that the mCherry (reporter expression; red) and GFP (reporter expression; green) overlaps with Ngn3 in the nucleus and E-cad at the membrane of the cells. (c) Construct of the Muc1mCherry reporter. (d) Representative section of a Muc1mCherry positive pancreas from an adult mouse, stained for Mucin1 (Muc1; green) shows that the mCherry (reporter expression; red) overlaps with Muc1 at the apical membrane. (e) Representative section of E15.5 WT pancreata (6 μm) stained for neurogenin3 (Ngn3; red), atypical protein kinase C (aPKC;green), mucin1 (Muc1; blue) and E-cadherin (Ecad; white). (f) Representative section of E15.5 WT pancreata (6 μm) stained for neurogenin3 (Ngn3; red), Crumbs3 (Crb3;green), mucin1 (Muc1; blue) and E-cadherin (Ecad; white). (g) Representative flow cytometry data from E12.5 Ngn3RG1 pancreata stained for Muc1, n = 3. (h) Quantification of the fraction of Ngn3 + cells that are Muc1 + (polarized) or Muc1- (Unpolarized) at E12.5, analyzed by Flow cytometry. n = 3 pancreata from each condition, from two litters, pTT < 0.0001. Error bars represent ± SEM. Scale bars, 20 μm. Statistical analysis: two-tailed student’s t-test. Statistic source data are found in Supplementary Table 3.
