Co-immunoprecipitation of full-length and truncated CaV2.2 is disrupted by the N-terminal construct.A: Representative Western blot showing proteins immunoprecipitated with rat anti-HA antibody and immunoblotted with anti-CaV2.2. tsA-201 cells expressed CaV2.2 HA, α2δ-1 and β1b, together with CaV3.1 Dom I–II (lane 1), CaV2.2 Dom I–II (lane 2), CaV2.2 Dom I–II plus GFP-CAAX (lane 3) or CaV2.2 Dom I–II plus CaV2.2-(43-95)-CAAX (lane 4). This result is representative of 3 independent experiments. B: Western-blot showing 30 μg WCL of the same samples shown in A. C: Quantification of Western blots. Band intensities for CaV2.2 Dom I–II were measured using ImageJ and normalised for expression of full-length CaV2.2 HA for each condition and relative to the condition of CaV2.2 HA + CaV2.2 Dom I–II (lane 2 in A and B). Data are mean band intensities (+ SEM) from three co-immunoprecipitation experiments from three different transfections. Statistical analysis: ns = non-significant difference, **p < 0.01.
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