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. 2004 Jan;123(1):53–62. doi: 10.1085/jgp.200308906

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Copyright © 2004, The Rockefeller University Press

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Figure 3. — Coexpression of wild-type and mutant CaM with TRPV1 alters the effects of Ca²⁺ on channel function in oocytes and HEK cells. (A) Box-plot of the percent decrease of capsaicin-activated (20 μM) current in Xenopus oocytes produced by 50 μM free Ca²⁺ and no added CaM (left, n = 8), 500 nM added CaM, (center, n = 8), or CaM_wt coexpressed with TRPV1 and no added CaM (right, n = 4). For the boxes, the line indicates the median of the data, the box surrounds the 25th through 75th percentile of the data, and the whiskers reach the 10th and 90th percentiles. As indicated by the asterisks, both the center and right groups had statistically significant differences compared with Ca²⁺ and no added CaM. (B) Box-plot of the percent decrease of capsaicin-activated (20 μM) current in HEK cells produced by 50 μM free Ca²⁺ and either no added CaM (first box, n = 7), 500 nM added CaM (second box, n = 3), CaM_wt coexpressed with TRPV1 and no added CaM (third box, n = 6), or CaM_1,2,3,4v. coexpressed with TRPV1 and no added CaM (fourth box, n = 6). As indicated by the asterisks, both the center groups had statistically significant differences compared with Ca²⁺ and no added CaM. For the fourth condition, p exactly equalled 0.05, and therefore did not meet our criterion for significance. (C) Effects of CaM_wt and CaM_1,2,3,₄ overexpression in HEK cells on the response to Ca²⁺ (50 μM). Open circles represent the data from coexpression of TRPV1 and CaM_wt and filled circles represent data from coexpression of TRPV1 and CaM_1,2,3,4. Data were obtained from pulses to +60 mV every 3 s. Ca²⁺ was introduced at time t = 0. (D) Dose–response to different Ca²⁺ concentrations in the presence of exogenous CaM. Data were obtained at a voltage of +60 mV and are normalized to the current obtained in the presence of each Ca²⁺ concentration with CaM. Data are the mean values from three patches. The smooth curve is a fit with the Hill equation, giving an IC₅₀ for inhibition by Ca²⁺ in the presence of CaM of 60 μM and a Hill slope of 1.25.