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. 2009 Jun;133(6):571–581. doi: 10.1085/jgp.200910206

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© 2009 Matsuba et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jgp.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 2. — Effects of PKA on cTnI phosphorylation and Ca²⁺ sensitivity in PLV reconstituted with cTn or PCRF. (A; Left) SDS-PAGE for control, cTn-, and PCRF-reconstituted PLV. Cont., control with no reconstitution; cTn, cTn reconstitution; PCRF, PCRF reconstitution; Tm, tropomyosin; LC-1, myosin light chain 1; LC-2, myosin light chain 2. (B) Western blotting showing the effect of PKA on cTnI Ser23/24 phosphorylation in cTn- and PCRF-reconstituted PLV (12.5 µg protein/well). Cont., control with no treatment. Normalized intensity compared with control (n = 3): cTn, 10.12 ± 6.53%; PCRF, 13.83 ± 5.23%; cTn + PKA (PKA phosphorylation after cTn reconstitution), 214.94 ± 30.82% (P < 0.05 compared with cTn); PCRF + PKA (PKA phosphorylation after PCRF reconstitution), 228.89 ± 20.13% (P < 0.05 compared with PCRF). (C) Force-pCa curves showing the effects of cTn reconstitution and subsequent PKA treatment on Ca²⁺ sensitivity. (Inset) Difference between the values of the PKA-induced shift of pCa₅₀ (C, control minus cTn + PKA; cTn, cTn minus cTn + PKA). *, P < 0.05. n = 7. (D) Force-pCa curves showing the effects of PCRF reconstitution and subsequent PKA treatment on Ca²⁺ sensitivity. (Inset) Difference between the values of the PKA-induced shift of pCa₅₀ (C, control minus PCRF + PKA; PCRF, PCRF minus PCRF + PKA). *, P < 0.05. n = 7. PKA treatment did not significantly affect maximal force or n_H in both C and D (see Table II).