Fig. 2: sNF96.2 cells secrete TGFβ and stimulate the SMAD signaling pathway in Capan-2 cells.
From: Schwann cells support oncogenic potential of pancreatic cancer cells through TGFβ signaling
a Amount of soluble TGFβ1 secreted into the medium by Capan-2 cells (Capan-2 CM) and by sNF96.2 cells (sNF96.2 CM). Quantification of soluble TGFβ1 concentration is represented as mean ± SD (n = 3 independent biological replicates, **P < 0.01). b Western blot detection of phospho-SMAD2 protein (P-SMAD2) in Capan-2 cells cultured for 1 h or 2 h either with Capan-2 CM or with sNF96.2 CM, and treated or not with the TβRI inhibitor, SB-431542. GAPDH and total SMAD2 were used as loading controls. Ratio of phospho-SMAD2 to total SMAD2 levels are indicated below the blots. c Immunofluorescence detection of SMAD2 protein (red) in Capan-2 cells cultured for 2 h alone (Capan-2/Capan-2 condition) or with sNF96.2 cells (sNF96.2/Capan-2 condition). Cell nuclei were counterstained with DAPI (blue). Scale bars, 30 μm. For each condition, images from one experiment representative of three independent experiment are shown (left panel) and quantification of nuclear SMAD2 per total number of nuclei is represented as mean ± SD (right panel, n = 3 independent experiments, *P < 0.05). CM conditioned medium.
