Fig. 3
From: ROBO2 is a stroma suppressor gene in the pancreas and acts via TGF-β signalling
Activation of Wnt and TGF-β pathway in cell cultures from Robo2F/F pancreas. a, b, c mRNA expression of Wnt targets (a), Tgfb1, Tgfbr2 (b) and Robo1 (c) analysed by RT-qPCR in D8 primary pancreatic cultures of Pdx1Cre and Robo2F/F animals. All qPCR data are referred to housekeeping gene Hprt. d Western blot (WB) analysis of phospho-Smad2 in D8 cultures and quantification of WB band density using ImageJ. e Representative images of mesenchymal cells, identified by Vim, in D8 Robo2F/F cultures stained with immunofluorescence/RISH for Axin2, Tgfb1, Robo1 or phospho-Smad2. Nuclei in immunofluorescence images are stained with DAPI. Images are representative of three to four independent experiments. Scale bars correspond to 20 µm. All data presented as mean + /− SEM; N = 4–6. Statistical analysis was performed using unpaired t test with Welch’s correction; *P < 0.05, **P < 0.01, ***P < 0.001
