Fig. 4

S-palmitoylation does not influence the total surface expression of HCN2. a Cell surface expression of ΔCysNTHCN2, the palmitoylation-deficient mutant of HCN2, examined using the cell surface biotinylation assay. The blots show biotin-labeled proteins probed with streptavidin-HRP (upper) and total proteins probed with anti-FLAG antibody and HRP-conjugated secondary antibody. b Effects of the S-palmitoylation inhibitor 2BP and the N-myristoylation inhibitor 2HM on the cell surface expression of HCN2 proteins. The transfected cells were incubated with 100 μM inhibitors for 24 h before the assay, as in (a). The signal intensities of biotin-labeled proteins were corrected for the expression level and normalized to the mean value of that of HCN2 (in a) or control (in b), and are shown below (n = 3). ns not significant versus HCN2 (in a) or control (in b)