APPENDIX
A. Adenovirus transfection with different gradients, the expressions of MG53/TGF-β1/Col - a1 and ZO-1 were detected by Western blot. The HNECs were transfected adenovirus for 72h. (When Adv-MOI =100, MG53 was significantly overexpressed and showed significant inhibitory effects on TGF-β1, Col-a1 and ZO-1.Therefore, Adv-MOI =100 was selected as the transfection condition).
B. The HNECs were transfected with adenovirus for 48 hours after transfection and photographed under fluorescence microscope(magnification 200X).
C. The HNCES were treated with different concentrations of TGF-β1 inhibitor for 72H.The inhibitory effect of MG53/TGF-β1/Smad/Col - a1 and ZO-1 were analyzed by Western blot. (When TGF-β1 inhibitor =50n M, the expression of MG53 /TGF-β1/ Col-a1 and ZO-1 were significant inhibited .Therefore, TGF-β1 inhibitor =50n M was selected as the optimum condition).