Fig. 3.

Immunodetection of Kv channel α subunits in cultured SCs. A, Cultured cells were subjected to immunofluorescence analysis with anti-S-100 antibody to confirm SC identity. Indirect immunofluorescence with anti-rabbit FITC was used for detection. Scale bar, 20 μm. B, Immunoblot analysis of Schwann cell cultures with antibodies to Kv channel α subunits. Membrane fractions of mouse primary cultured SCs were subjected to immunoblot analysis with subunit-specific antibodies to Kv1.1, Kv1.2, Kv1.4, Kv1.5, Kv2.1, Kv3.1b, and Kv3.2 α subunits. To check the specificity of antibody labeling toward SC membrane extracts, each antiserum was preincubated for 1 hr at room temperature in the presence (+) or absence (-) of its respective antigen, as indicated in Materials and Methods. HRP-conjugated secondary antibodies and ECL were used for detection.