Fig. 3

EDAR PM trafficking requires PKA activation and SNAP23. A–B Immunoblotting shows the intensity of phosphorylated PKA substrates (PKA activity) with or without treatment of EDA, H89 (60 μM), or Forskolin (20 μM). C Immunoblotting shows the levels of indicated proteins in Cyt or PM fraction with or without treatment of EDA, H89 or Forskolin. D The localization of EDAR-GFP in EDAR-GFP⁺ cells with or without indicated treatment of EDA, H89 or Forskolin. E Quantification shows the ratio of EDAR levels in PM to Cyt in D. Data from at least 10 random images in each condition from 3 independent experiments. F As in (D), except with indicated treatment of EDA and transfection of SNAP23 siRNAs. G As in C, except with indicated treatment of EDA and transfection of SNAP23 siRNAs. Error bars indicate mean ± SD from at least 30 cells from 3 independent experiments. ***P < 0.001; Student’s t-test. Scale bar, 20 μm