EphrinA2 Regulates Clathrin Mediated KSHV Endocytosis in Fibroblast Cells by Coordinating Integrin-Associated Signaling and c-Cbl Directed Polyubiquitination
Figure 5
EphA2 associated with integrins and clathrin are localized to non-lipid rafts in KSHV infected HFF cells.
(A) Serum-starved HFF cells were either uninfected or infected with KSHV (30 DNA copies/cell) for the indicated time points. LR and non-LR fractions were isolated and analyzed by Western blotting for the presence of EphA2. Caveolin-1 and CD71 characterize the purity of LR and non-LR fractions, respectively. (B) Western immunoblot analysis showing phosphorylation of EphA2 at the indicated time post-KSHV infection in non-NLR fractions from HFF cells. (C) 150 µg of protein from non-LR fractions of uninfected and KSHV-infected cells was immunoprecipitated with anti- αVβ5 or anti-EphA2 antibodies at 4°C overnight followed by Western blotting with EphA2 or clathrin heavy chain, respectively. Blots were stripped and reprobed for β5 and β3, respectively. CD71 characterizes the purity of non-LR fractions.