Fig. 6.

CaM₁₂ enhances the total expression level of Ca_V2.2. A. Representative Western blot of Ca_V2.2 in total lysates. Cell lysates isolated from HEK293T cells with no transfection (None), transfected with T-Ca_V2.2 only (T-Ca_V2.2, Mock), or transfected with co-expression of CaM, CaM₁₂, CaM₃₄, and CaM₁₂₃₄ were used for Western blot analysis using antibodies against FLAG and β-actin. B. Normalized total Ca_V2.2 level. The level of T-Ca_V2.2 was normalized to the level of β-actin in each sample; the values were then normalized to the value of the CaM group in each independent experiment. C. Representative staining of Ca_V2.2 after being biotinylated. Membrane proteins were first labeled with biotin and then isolated with avidin. The T-Ca_V2.2 in the total lysate (Total) and avidin-purified fraction (Biotinylated) was analyzed by an antibody against FLAG. D. The ratio of Ca_V2.2 in the membrane fraction to the total lysate. Data are presented as the mean ± SEM from at least 3 independent experiments and were analyzed by one-way ANOVA with Fisher's post hoc test. *: p < 0.05 compared to the CaM group.