Journal: Cells

Article Title: Proteomics Approach Highlights Early Changes in Human Fibroblasts-Pancreatic Ductal Adenocarcinoma Cells Crosstalk

doi: 10.3390/cells11071160

Figure Lengend Snippet: BAG3 is strongly induced in HDF cells treated with MIA-PaCa-2 CM. ( A ) PPI network predicted by STRING among the proteins identified in lysate of HDF cells treated for 48 h with CM. BAG3 is highlighted by a red circle. ( B ) Western blot of BAG3 in HDF cell lysate treated for 24 h and 48 h with MIA-PaCa-2 CM.

Article Snippet: Binding of non-specific antibodies was obtained by blocking the membrane with 5% not-fat dry milk in PBS with 0.01% Tween 20 for 1 h at RT and then incubated overnight with the following antibodies: anti-p44/42 MAPK (Erk1/2, MAPK3/1, 1:2000), anti-SMAD2/3 (1:1000), anti-GAPDH (1:1000) and anti-STAT3 (1:1000) (Cell Signaling Technology, Danvers, MA, USA); anti-LDHA (1:1000) and anti-alpha-SMA (1:500) (Thermo Scientific, Rockford, IL, USA); anti-ITGB3 used 1:1000 (Abcam, Cambridge, UK), anti-BAG3 used 1:2000 (Novus Biologicals, Milan, Italy) and anti-β-actin used 1:40000 (Sigma-Aldrich, Milan, Italy).

Techniques: Western Blot