FIGURE 5.

Induction of CaR^T888 phosphorylation by the positive allosteric CaR modulators l-Phe and NPS-R467. A, representative immunoblots showing CaR^pT888 (i) and total CaR (ii) immunoreactivity in CaR-HEK cells following treatment with 1.2 mm Ca²⁺_o in the presence or absence of 1 μm NPS-R467 or 10 mm l-Phe for 2, 5, and 10 min. Quantification of the changes in 160 kDa CaR^pT888 immunoreactivity is shown in panel iii (n = 9). *, p < 0.05, **, p < 0.01 (R-467), +p < 0.05 (l-Phe) versus 1.2 mm Ca²⁺_o by one-way ANOVA (Dunnett's post test). Representative traces showing Ca²⁺_i changes in a single cell (gray) or global cluster of cells (black) in response to 1 μm NPS-R467 (Bi) or 10 mm l-Phe (Ci) (in the presence of 1.5 mm Ca²⁺_o). Quantification of the Ca²⁺_i changes is shown as area under the curve/min in panels Bii and Cii (n = 3). *, p < 0.05, ***, p < 0.001 versus responses before and after allosteric treatments, by repeated measures ANOVA. arb., arbitrary; Cont, control.