Fig. 2
From: Ankyrin B and Ankyrin B variants differentially modulate intracellular and surface Cav2.1 levels
Wildtype AnkB, AnkB p.S646F and AnkB p.Q879R do not impact Cav2.1 surface expression levels. HEK293T cells were co-transfected with Cav2.1, and GFP control, wildtype, or mutant AnkB-GFP. (ai) Representative surface biotinylation Western blots of inputs and NeutrAvidin surface fraction. GAPDH was used as a negative control to ensure intracellular proteins were not biotinylated. Transferrin receptor (TfR) served as a positive pulldown control, (aii) 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 = 3–4, F (2, 8) = 1.222, p = 0.3443 (Cav2.1: WT,S646F); N = 3, F (2, 6) = 5.891, p = 0.0384 (Cav2.1: Q879R, E1458G); N = 3–4, F (2, 8) = 5.764, p = 0.0282 (GFP: WT, S646F); N = 3, F (2, 6) = 2.393, p = 0.1721 (GFP: Q879R, E1458G). This data is included in the MSc thesis of CSWC, University of Victoria, 2019 found at https://dspace.library.uvic.ca//handle/1828/11053