Effect of EphA2 knockdown on the activation of KSHV induced signalling molecules as well as endocytic effectors.

<p>(<b>A</b>) Control or EphA2 shRNA-transduced HFF cells were either mock infected (UI) or infected with KSHV (30 DNA copies/cell) for the indicated time period and subjected to Western blot analysis for phospho-FAK (p-FAK) (Y397), phospho-Src (Y-416) and phospho-PI3-K (Y458). Blots were stripped and reprobed for the respective total FAK, Src and PI3-K. Efficiency of EphA2 knockdown was analyzed by Western blot with EphA2 antibody and β-actin was used as loading control. (<b>B</b>) Control or EphA2 shRNA transduced HFF cells either mock infected or infected with KSHV (30 DNA copies/cell) were subjected to Western blot analysis for activation (phosphorylation) of the indicated endocytic effector molecules (c-Cbl, myosin IIA and clathrin). For clathrin activation, cell lysates were subjected to immunoprecipitation with clathrin heavy chain antibody followed by Western blot analysis with anti-phosphotyrosine antibody (4G10). Blots were reprobed for the respective total c-Cbl, myosin IIA and clathrin for equal expression. The levels of fold activation are indicated.