Fig. 3
From: Ankyrin B and Ankyrin B variants differentially modulate intracellular and surface Cav2.1 levels
AnkB and its variants differentially impact Cav2.1 expression levels in the presence of auxiliary subunits. HEK293T cells co-transfected with Cav2.1, α2δ1, β4, and GFP control, wildtype, or mutant AnkB-GFP (ai) Representative surface biotinylation Western blots of inputs and surface fraction. (aii) Quantification of overall input proteins normalized to GAPDH and expressed as a percentage of GFP control (CTL). One-way ANOVA followed by Dunnett’s multiple comparison, N = 6, F (2, 15) = 6.072, p = 0.0117 (Cav2.1: WT,S646F); N = 6, F (2, 15) = 0.5018, p = 0.6152 (Cav2.1: Q879R, E1458G); N = 6, F (2, 15) = 0.8088, p = 0.4639 (GFP: WT,S646F); N = 6, F (2, 15) = 36.21, p < 0.0001 (GFP: Q879R, E1458G); N = 6, F (2, 15) = 125.6, p < 0.0001 (α2δ1: WT,S646F); N = 6, F (2, 15) = 32.05, p < 0.0001 (α2δ1: Q879R, E1458G); N = 6, F (2, 15) = 90.86, p < 0.0001 (β4: WT,S646F); N = 6, F (2, 15) = 30.48, p < 0.0001 (β4: Q879R, E1458G). (aiii) Quantification of surface proteins normalized to surface TfR and expressed as a percentage of GFP control (CTL). One-way ANOVA followed by Dunnett’s multiple comparison, N = 3, F (2, 6) = 0.1117, p = 0.8961 (Cav2.1: WT,S646F); N = 3, F (2, 6) = 25.14, p = 0.0012 (Cav2.1: Q879R, E1458G); N = 3, F (2, 6) = 59.18, p = 0.0001 (GFP: WT,S646F); N = 3, F (2, 6) = 9.587, p = 0.0135 (GFP: Q879R, E1458G); N = 3, F (2, 6) = 14.18, p = 0.0053 (α2δ1: WT,S646F); N = 3, F (2, 6) = 0.6860, p = 0.5392 (α2δ1: Q879R, E1458G). A portion of this data is included in the MSc thesis of CSWC, University of Victoria, 2019 found at https://dspace.library.uvic.ca//handle/1828/11053