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. Author manuscript; available in PMC: 2010 Jul 24.
Published in final edited form as: Nat Chem Biol. 2007 Jun 17;3(7):423–431. doi: 10.1038/nchembio.2007.4

Figure 3.

Figure 3

Ca2+ transients in response to three 20-ms depolarization pulses. (a) A schematic representation of an α1C channel tagged with GFP and the optimized tetracysteine peptide N4 (N4-GFP-α1C). (b) A TIRF image showing the locations of the color-coded and numbered ROIs. The black ROI is the entire cell (not shown). (c) Current trace with three short depolarization pulses. Time is measured from the beginning of the first pulse. 1 mM BAPTA was present in the pipette; the external solution contains 15 mM Ca2+ and 10 μM FPL 64176. (d) Three fluorescent traces aligned in time with the current trace. The imaging rate was 100 f.p.s. (e) Two additional fluorescent traces registered at ROIs placed on hot spots. (f) A second current applied to the same cell, 2 min after the trace shown in c, using the protocol of Figure 2d. (g) Fluorescent signals at the ROI shown in c in response to the 960-ms depolarization. (h) Comparison of the responses of the red ROI to the two protocols, depicted as F/F0 (the color of the trace from d was switched to gray).