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. 2013 May;141(5):585–600. doi: 10.1085/jgp.201210861

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© 2013 Grubb et al.

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Figure 4. — Ca²⁺ activation kinetics of mTMEM16F. (A) Typical current-voltage relationships of mTMEM16F at max current with 90, 190, or 250 µM free [Ca²⁺]_i. (B) The mean mTMEM16F I-V relation at maximal activation with the indicated free [Ca²⁺]_i, generated from ramp protocol sweeps (from −120 to 80 mV). Data are presented as mean ± SE of three to nine replicates. (C) Hill plot of normalized mTMEM16F I_Cl,Ca densities at 70 mV. Data are presented as mean ± SE of 3–12 replicates. (D) Ca²⁺ dependence of mTMEM16F versus time from whole start of whole-cell recording to peak I_Ca,Cl activation. Linear regression is included to illustrate the decline in the time to peak activation with increasing free [Ca²⁺]_i and is given by f(x) = −0.04x + 14.11. (E) mTMEM16F-GFP expressed in HEK293 cells. Immediately before fixation, the cells were exposed to ionomycin for 20 min in a solution containing 500 µM free Ca²⁺. The images were taken using an epifluorescence microscope. mTMEM16F-GFP was visualized with anti-GFP and Alexa Fluor 488, whereas the plasma membrane was stained using rhodamine-conjugated WGA. The nucleus was stained with DAPI.