Fig. 4
From: Novel TRPV6 mutations in the spectrum of transient neonatal hyperparathyroidism
Western blot analysis of TRPV6 mutants. a Localization of TRPV6 proteins. Plasma membrane proteins in TRPV6-transfected HEK293T cells were biotinylated and collected with streptavidin beads. Myc-TRPV6 proteins were analyzed by western blotting using anti c-myc antibody (left). An ~ 80-kDa band was observed for cells expressing WT TRPV6 but not the p.Arg390His or p.Gly428Arg mutants. Bands for WT, p.Arg390His, and p.Gly428Arg were observed in total lysate (right), suggesting that the mutants had impaired trafficking to the plasma membrane. b Conventional western blotting of total cell lysates. Signals for p.Gly291Ser and p.Gly451Glu were significantly lower than that for WT in cells not treated with ruthenium red. In the presence of ruthenium red (4 µM) the signals were recovered, suggesting that cells expressing the mutants had Ca2+ overload