Figure 3: Tgif2 expression induces adult murine liver cells to acquire molecular features of pancreatic progenitors.

(a) Map of LVs and schematic of the ex vivo reprogramming. Murine adult primary or BAML hepatocytes (HEP) were transduced with the constitutively active PGK-TGIF2-EGFP LV (LV-TGIF2) expressing Tgif2 and EGFP, FACS-sorted for EGFP and characterized at different time points by various ex vivo and in vivo approaches. (b) RT-qPCR analysis of hepatic and pancreatic gene expression in mouse primary HEPs at d7 and d14 after transduction with LV-TGIF2. Data were normalized to Sdha and represented as Log2-expression ratio between LV-TGIF2-transduced and control hepatocytes at matched time-points. (c) RT-qPCR analysis of hepatic and pancreatic gene expression in murine adult BAML HEP cells transduced with LV-TGIF2 at the indicated time points after transduction. Data were normalized to Sdha and represented as Log2-expression ratio between LV-TGIF2-transduced and control cells. Values shown are mean±s.e.m. (n=5) *P<0.05. (d) IF of unsorted (no FACS) LV-TGIF2-transduced (right) BAML HEP cells and control cells (left). In top panel of LV-TGIF2-transduced cells, asterisks (*) indicate Albumin (ALB)-positive cells that are PDX1-negative; open arrowheads (>) indicate PDX1/GFP-positive cells that are ALB-negative in transduced cultures. In middle panels, arrows indicate examples of GFP-positive cells either PDX/SOX9-double or PDX1/TGIF2-double positive cells. In bottom panel, asterisks (*) indicate Glutamine synthetase (GS)-positive cells (blue) that are negative for PDX1 (red) in cultures d50 after transduction; open arrowheads (>) indicate PDX1-positive cells that are negative for GS. Scale bars, 50 μm.