Fig. 6

T346M blocks EDAR transportation and completely restrains MG growth. A Fluorescence images show the location of EDAR-T346M expressed by AAV particles also encoding GFP separated by T2A motif. B LEF1 (red) and K14 (green) IHC staining of cultured WT eyelids transduced by AAV particles encoding GFP, EDAR or T346M. C Quantitation of MG length in B. D GFP-IP was conducted using cytoplasmic lysates from cells treated with or without EDA and expression of WT, EDAR or T346M. Immunoblotting shows the protein levels using the indicated antibodies. Red rectangle shows SNAP23 with obvious changes. E Immunoblotting shows the intensity of PKA activity in cells expressing T346M with or without EDA treatment. F Immunoblotting images show the levels of indicated proteins in Cyt and PM fractions from cells expressing T346M with or without EDA treatment. Error bars indicate mean ± SD from at least 20 MGs in total 3 independent experiments. ***P < 0.001; Student’s t-test. Scale bar, 20 μm