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Fig. 8. Inhibition of Ca²⁺-dependent nuclear translocation of 6ROX-CaM by mTrp peptide in HeLa cells. (A,B) Fluorescence of (A) 6ROX-CaM and (B) fluo-4 in HeLa cells in 2 mM EGTA incubated with 10 µM mTrp-peptide for 15 minutes (t₀). At t₅₀ cells were stimulated with 2.1 mM Ca²⁺. (C,D) Time course of normalized fluorescence (F/F_(O)cytoplasm) of 6ROX-CaM (C) and normalized fluorescence (F/F_(O)cytoplasm) of fluo-4 (D) in the nucleolus (), nucleoplasm () and cytoplasm (). Arrow in C indicates the addition of 2.1 mM Ca²⁺. (E) Relative fluorescence of TA-CaM with increasing concentrations of AIP. A solution of 21 nM TA-CaM, 100 mM KCl, 2 mM MgCl₂ and 50 mM K-MES, pH 7.0 at 21°C was titrated with up to 140 nM mAIP in 100 µM Ca²⁺ (). (F) 6ROX-CaM diffusion rate at elevated levels of intracellular Ca²⁺. HeLa cells were electroporated with 6ROX-CaM and stimulated with 2.1 mM Ca²⁺ in the presence of 10 µM mTrp peptide for 30 minutes. The F_np/F_cytoplasm ratio over a period of 30 minutes was monitored.

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