Co-immunoprecipitation of SNAP23 with GLUT1, Plin2, or Plin1.

<p>The native proteins in cell homogenates from Plin2 overexpressing and control cells were co-immunoprecipitated as described in the Methods section. To examine SNAP23/GLUT1 interactions, GLUT1 was immunoprecipitated using anti-GLUT1. Levels of SNAP23 (A) in the immunoprecipitate from control (lane 1) and PLIN2 overexpressing cells (lane 2) were analyzed by immunoblotting with anti-SNAP23. Equal immunoprecipitation of GLUT1 was verified by immunoblotting with anti-GLUT1 (B). The ratio of SNAP23 immunoprecipitated with GLUT1 was calculated from the integrated density values from Western blots (C). To analyze SNAP23/Plin2 interactions, Plin2 was immunoprecipitated with anti-Plin2 and levels of SNAP23 (D) were detected by immunoblotting with anti-SNAP23. Equal immunoprecipitation of Plin2 was verified by immunoblotting with anti-Plin2 (E). The ratio of SNAP23 immunoprecipitated with GLUT1 was calculated from the integrated density values from Western blots (F). Reverse immunoprecipitation experiments were also performed. SNAP23 was immunoprecipitated with anti-SNAP23 and levels of Plin2 (G), GLUT1 (H), and Plin1 (I) in the immunoprecipitate were analyzed by Western blotting. Equal immunoprecipitation of SNAP23 was verified by immunoblotting (J). The ratio of Plin2, GLUT1 and Plin1 immunoprecipitated with SNAP23 was calculated from the integrated density values from Western blots (K). Immunoprecipitate obtained using secondary IgG antibodies (lane 3) were used as negative controls for each set. (*) indicates p < 0.05 as compared to control.