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Fig. 5. Co-existence of MIP(AQP0) in the complex formed by Cx45.6/Cx56. Gap-junction-rich embryonic lens membranes were isolated from embryonic-day-11 lenses and immunoprecipitated with immobilized affinity-purified anti-Cx45.6 (A-C, lanes 1 and 2), anti-Cx56 antibodies (A-C, lanes 3 and 4) or anti-MIP(AQP0) antiserum (A-C, lanes 5 and 6) in the absence (A-C, lanes 1, 3 and 5) or presence (A-C, lanes 2, 4 and 6) of SDS. The resulting immunoprecipitates were immunoblotted with affinity-purified anti-Cx45.6 (A) or anti-Cx56 (B) antibodies, or with anti-MIP(AQP0) antiserum (C). The SDS-boiled precipitates of Cx45.6-antibody-conjugated beads alone were blotted by anti-MIP(AQP0) antiserum (C, lane 7). The cross-reacting immunoglobulin light chain was shown as (*). (D) Silver staining of the samples from membrane preparation of embryonic-day-10 lens (lane 1), supernatant of immunoprecipitation of lens membranes with anti-Cx45.6 antibody (lane 2), immunoprecipitates of lens membrane with anti-Cx45.6 antibody (lane 3), fivefold overloaded sample (lane 4), SDS-boiled precipitates with Cx45.6-antibody-conjugated beads in the absence of lysates (lane 5) and precipitates with non-Cx45.6-antibody-conjugated beads (lane 6) in the presence of lysates. The immunoprecipitates of lens membranes with anti-Cx45.6 antibody were immunoblotted by anti-MIP(AQP0) monoclonal antibody (lane 7).

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