Fig. 2.
CCN5 Binds Integrin αvβ3. a Exponentially growing VSMC were infected with either adenovirus expressing an HA tagged CCN5 (lane 1) or adenovirus expressing GFP (lane 2), after which cell lysates were prepared in an NP-40 based lysis buffer. Equal amounts of lysate protein were immunoprecipitated using anti-HA bound magnetic beads. Bound proteins were resolved by SDS-PAGE and analyzed by Western blotting using an antibody against Integrin β3, Integrin αv, Integrin αIIB (as a control for nonspecific integrin binding), or HA (to demonstrate HA-CCN5 binding to beads). b VSMC were growth arrested by serum starvation, after which, cell lysates were prepared in an NP-40 based lysis buffer. Equal amounts of lysate protein were immunoprecipitated using magnetic beads bound to either antibody against Integrin β3 (lane 1) or mixed IgG (lane 2). Bound proteins were resolved by SDS-PAGE and analyzed by Western blotting using an antibody against CCN5, or Integrin β3 (to demonstrate binding of Integrin β3 to beads). Both experiments replicated three times