Figure 3.

Biochemical and Cellular Analysis of mLoF and mGoF CaSR Mutations

(A) Plot presents mean ± SD of western blot analysis of lysates from three independent HEK293 cell transfections, as the percent of mature receptors ([mature/(immature + mature)]%) for WT (black bar), mGoF (white bars), and mLoF (gray bars) variants. Representative western blots presented in Figure S3. Statistical analysis was by Student’s t test, with ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, and ^∗∗∗∗p < 0.0001.

(B) Plasma membrane localization was determined for WT (black bar), mGoF (white bars), and mLoF (gray bars) variants by ELISA of transiently transfected HEK293 cells, plotted as the mean ± SD of the percent of surface to total expression, for at least three independent experiments. Statistical analysis was by Student’s t test, with ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, and ^∗∗∗∗p < 0.0001.

(C) Bar graph of EC₅₀ values for WT (black bar), mGoF (white bars), and mLoF (gray bars) variants extracted from fits of dose-response relations for Ca²⁺_e. Statistical analysis by F-test, with ^∗∗∗∗p < 0.0001. Titrations were done as described in Subjects and Methods, and plots of dose-response relations which were fitted to extract EC₅₀s are presented in Figures S5 and S6. Numbering of residues for all graphs based on GenBank: NM_000388.4.