Figure 7: In vivo analysis of TGIF2 reprogramming activity.

(a) Schematic of the AAV-mediated experimental strategy to express Tgif2 in vivo in the adult mouse liver. AAV-injected and uninjected control livers were examined at the indicated time points (*) after iv injection. (b) RT-qPCR analysis of indicated genes in AAV.TGIF2-injected livers at d30. Data were normalized to 36B4 and represented as Log2-expression ratio between AAV.TGIF2-injected and AAV.GFP-injected livers. Values shown are mean±s.e.m. (n=3). (c) IF staining of AAV-injected (+AAV.TGIF2) and uninjected (−AAV) adult mouse livers for GS, SOX9, Epcam, PDX1 and NKX6.1. Micrographs show IF stainings performed at d30 after AAV.TGIF2 injection; SOX9/NKX6.1 IF staining was done at d60. SOX9 and Epcam mark biliary epithelial cells (bec) and are absent in hepatocytes in uninjected livers. Arrows indicate examples of SOX9−, PDX1− or NKX6.1-positive hepatocytes in AAV.TGIF2-injected livers. Outlined areas are shown in the insets without Hoechst nuclear counterstain (blue). CV, central vein. Scale bars, 50 μm. (d) Quantification of SOX9-positive hepatocytes and PDX1-positive cells in adult livers injected with AAV.GFP (n=2 d30), AAV.TGIF2 (n=3 d7; n=4 d30) and uninjected (n=3). Average number of labelled cells per section (mm²) was determined for each animal in one entire liver lobe. Results are expressed as the mean±s.e.m. Statistics by two-tailed t-test. **P<0.01.