Fig. 5.

Lineage tracing of Pdx1-expressing progenitors in Robo1/2 KO embryos. a Schematic representation of the transgenes and breeding scheme used for lineage tracing experiments in Robo1/2 KO embryos. b Quantification of GFP⁺ cells in the E12.5 liver of CTRL (n = 3) and Robo1/2 KO (n = 4) embryos. Number of GFP⁺ cells was normalised to the liver bud volume. Error bars represent ± s.e.m. Two-tailed Student’s t-test *P < 0.05. c Representative tiled IF image of E12.5 Pdx1-Cre;H2B-GFP;Robo1^+/+/Robo2^+/+ (CTRL) cryosections stained for GFP and Hoechst (Hoe) nuclear counterstain. As expected, cells in ventral pancreas (vp), dorsal pancreas (dp), stomach (st) and duodenum were labelled by Pdx1-Cre^24,29. Scale bars, 200 µm. d Representative IF images of E12.5 and E14.5 Pdx1-Cre;H2B-GFP;Robo1/2 KO (Robo1/2 KO) cryosections stained with indicated antibodies and Hoechst (Hoe) as nuclear counterstain. Arrowheads indicate GFP⁺ cells in the liver (lv), which are also Prox1⁺ or Albumin⁺ (Alb). e Representative IF images of E18.5 Pdx1-Cre;H2B-GFP;Robo1/2 KO (Robo1/2 KO) cryosections stained with indicated antibodies and Hoechst (Hoe) as nuclear counterstain. Arrowheads indicate GFP/Albumin (Alb)-double positive cells in the hepatic parenchyma and GFP/CK19-double positive cells in the portal vein (PV) space. Scale bars, 50 µm