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. 2017 Feb 21;66(5):1293–1300. doi: 10.2337/db16-0887

Figure 1.

Figure 1

Mafa and Pdx1 enhance production of insulin-positive cells from glucagon-positive cells when coexpressed in pancreatic endocrine precursor cells. A: Schematic representation of Mafamyc, Pdx1flag, and Cre-mediated recombination. Exogenous myc-tagged Mafa or flag-tagged Pdx1 is expressed via the excision of the stuffer chloramphenicol acetyltransferase (CAT) cassette upon production in islet cell precursors of Ngn3-Cre or of Gcg-Cre in islet α-cells. B: MafA and Pdx1 antibodies were used to show ectopically expressed Mafamyc and Pdx1flag in essentially all Ngn3-Cre;CAT-Mafa (left) or Ngn3-Cre;CAT-Pdx1 (right) islet glucagon-positive cells at P0.5. The arrows illustrate Mafa or Pdx1 expressing glucagon-positive cells. C: Insulin-positive and glucagon-positive cells in representative islets of the Ngn3-Cre, Ngn3-Cre;CAT-Mafa, Ngn3-Cre;CAT-Pdx1, and Ngn3-Cre;CAT-Mafa;CAT-Pdx1 pancreas at P0.5 or 6 weeks. The arrows depict insulin and glucagon–copositive cells, which are only detected in the Ngn3-Cre;CAT-Mafa;CAT-Pdx1 pancreas at P0.5. D: Quantitative analysis of the islet insulin-, glucagon-, or cohormone-positive production at P0.5 and 6-week-old cells in the Ngn3-Cre, Ngn3-Cre;CAT-Mafa, Ngn3-Cre;CAT-Pdx1, and Ngn3-Cre;CAT-Mafa;CAT-Pdx1 pancreas. The ratio of the insulin-, glucagon-, or double-positive cells is presented relative to the total hormone-positive cell number. The ratio of the double hormone–positive cells to glucagon-positive cells was compared at P0.5 and of glucagon-positive cells to insulin-positive cells at 6 weeks. n = 3–6. 6W, six weeks; CAG, chicken β-actin promoter and cytomegalovirus enhancer; INS, insulin. *P < 0.001. Scale bars: 50 μm.