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. Author manuscript; available in PMC: 2011 Jul 1.
Published in final edited form as: Nat Cell Biol. 2010 Dec 19;13(1):49–58. doi: 10.1038/ncb2133

Figure 3. DR1 interacts with Par3 and Par 6.

Figure 3

(a) Comparison of the last C-terminal aminoacids of DDR1 in human, rat, mouse, claudin 4, 5 and 6 and DDR2. (b) Negative staining of GST pulldown using GST alone as control (left), DDR1 amino acids 801-876 (middle) and DDR1 amino acids 801-875 (right). Asterisks show bands present only in the 801-876 lane. Bottom panel: Par3 western blot showing Par3 pulled down only by DDR1 801-876. (c) i) Non-transfected (first lane), GFP-transfected (second lane) or Par3-GFP-transfected (third lane) A431 cell lysates were incubated with anti-GFP antibody and the amount of endogenous DDR1 bound was determined by western blot. The Par3 western blot is shown on the middle. The amount of DDR1 in the starting lysates is shown on the bottom. ii) Non-transfected (first lane), empty PRK5.1-transfected (second lane) or Par6-flag-transfected (third lane) A431 cell lysates were incubated with anti-Flag antibody and the amount of endogenous DDR1 bound was determined by western blot. The amount of DDR1 in the starting lysates is shown on the bottom. (d) Flag-tagged PDZ domains of Par3 (third to fifth lanes) and Par6 (sixth lane) were immunoprecipitated from 293 cells also expressing DDR1. Non-transfected cells (first lane) and DDR1 alone (second lane) were used as controls. (e) Representative pictures showing Par3, DDR1 and F-actin staining (blue in merge) in A431 cells in apical membranes. Scale bar is 10 μm. (f) representative pictures showing actin (red) and Par3 (green) localisation in two clones stably knocked down of DDR1 (shDDR1#3 or shDDR1#4) or control A431 cells (shCtr) Scale bars are 10μm. (g) Representative pictures of Par3 (top) and DDR1 (bottom) in A431 cells transfected with control siRNA (left panels) or siRNA against Par3 (right panels). Scale bar is 20 μm.