Fig. 7.

Par3-S383A and Par3-S383D show different patterns of subcellular localization. (A) Hippocampal neurons were transfected at 0 DIV with vectors encoding GFP and Myc–Par3-S383A or Myc–Par3-S383D. Neurons were fixed at 3 DIV and stained with an anti-Myc antibody (Par3). A higher magnification of the areas marked by squares is shown below each image. Scale bars: 50 µm. (B) The relative intensity of the immunofluorescence in the axon (gray bars) or soma and dendrites (black bars) for staining with the anti-phospho-Ser383-Par3 antibody was quantified as the percentage of the total signal for phospho-Ser383-Par3. Values are the mean±s.e.m. (n = 6); **P<0.01 between indicated pairs; two-way ANOVA. (C) RFP–Dlic2 (DIC) or RFP (negative control) were coexpressed with Myc-tagged wild-type (WT) Par3, phospho-mimic Par3-S383D or non-phosphorylatable Par3-S383A in HEK 293T cells. Dlic2 was immunoprecipitated (IP) from cell lysates with an anti-RFP antibody, and the bound proteins were detected by western blotting (WB) using an anti-Myc antibody. Blotting for RFP, Myc and tubulin (input) confirmed that comparable amounts of proteins were expressed and loaded. Numbers indicate the molecular mass in kDa.