Figure 3
From: Rho GTPase Inactivation Impairs Lens Growth and Integrity
A, Detection of C3 protein by Western blot analysis. Although homogenates obtained from wild-type lens (lane 1) showed no immunoreactivity to C3 monoclonal antibody, lens homogenates obtained from line ODO4-11 (lane 2) and line ODO4-21 (lane 3) C3 mice showed a immunopositive band (≈25 kDa). B, To test the functional integrity of the C3 protein, lens homogenates from transgenic and wild-type littermates were subjected to in vitro ADP-ribosylation of Rho GTPase by exogenous C3-exoenzyme. Lane 1 is a control in which the C3-exoenzyme was absent. As compared with a wild-type sample (lane 2), transgenic lens homogenate obtained from line ODO4-21 mice showed a very weak in vitro ADP-ribosylated band (lane 3), indicating that most of the Rho GTPase had already been ribosylated by the endogenous C3 produced transgenically, suggesting its in vivo inactivation. The lower dark band is derived from the unbound radiolabeled compound.
