Figure 6: In vivo analysis of TGIF2 reprogramming activity.

(a) Transplantation assays in Akita diabetic mice. Reprogrammed TiPP cells (derived from BAML HEPs+LV-TGIF2 d8 post-transduction) were grafted under the KC of hyperglycemic Akita mice (red line). Animals engrafted with BAML HEP cells transduced with LV-GFP were used as negative controls (green line). Blood glucose levels were measured under non-fasting conditions before transplantation (time points: −2 wk and −1 wk), the day of transplantation (time point 0; arrow) and, subsequently, twice a week after transplantation. n=6 animals in each group. Normoglycemia is defined as blood glucose level below 200 mg dl⁻¹ in wild-type littermate mice under nonfasting conditions (dotted line). Values shown are mean±s.e.m. **P<0.01 (two-way ANOVA comparison test between LV-GFP and TiPP transplanted groups). (b) IF analysis on cryosections of mouse KCs transplanted with LV-GFP BAML HEP cells and TiPP cells. Hoechst (Hoe) nuclear counterstain in blue. Insets show area in the dashed box of: PDX1 (red)/GFP (green) staining without SOX9 channel (blue); C-peptide (C-PEP) (red)/Hoechst (blue) staining without GFP channel (green); PC2 (red)/Hoechst (blue) staining without GFP channel (green); UCN3 (red)/Hoechst (blue) staining without GFP channel (green); GLUT2 (red)/Hoechst (blue) staining without GFP channel (green), arrows indicate staining near the membrane, as expected for the location of mature Glut2 protein. To a lesser extent, some cells displayed Glut2 staining in the cytoplasm, which is similar to neonatal beta-cells. Scale bars, 50 μm.