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(A-C) show myofilament Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation compared to donor tissue. (D-F) show Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation before and after normalizing <t>cTnI</t> phosphorylation by incubation with protein kinase A (PKA). (G) and (H) display the phosphorylation status of cardiac troponin I determined by phos-tag gel analysis before and after incubation with PKA. 0P <t>represents</t> <t>non-phosphorylated</t> cTnI, 1P single-phosphorylated cTnI and 2P double-phosphorylated cTnI. (G) shows the phos-tag gel image for the quantified data in (H). (I-K) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 compared to donor. (L-N) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 before and after PKA treatment. * p < 0.05, ** p < 0.01, unpaired t -test. n(Donor) = 4, n(HCM 173) = 4/3, n(HCM 175) = 4/3, R278C = 3/3 for with/without PKA, respectively.
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(A-C) show myofilament Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation compared to donor tissue. (D-F) show Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation before and after normalizing <t>cTnI</t> phosphorylation by incubation with protein kinase A (PKA). (G) and (H) display the phosphorylation status of cardiac troponin I determined by phos-tag gel analysis before and after incubation with PKA. 0P <t>represents</t> <t>non-phosphorylated</t> cTnI, 1P single-phosphorylated cTnI and 2P double-phosphorylated cTnI. (G) shows the phos-tag gel image for the quantified data in (H). (I-K) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 compared to donor. (L-N) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 before and after PKA treatment. * p < 0.05, ** p < 0.01, unpaired t -test. n(Donor) = 4, n(HCM 173) = 4/3, n(HCM 175) = 4/3, R278C = 3/3 for with/without PKA, respectively.
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(A-C) show myofilament Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation compared to donor tissue. (D-F) show Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation before and after normalizing <t>cTnI</t> phosphorylation by incubation with protein kinase A (PKA). (G) and (H) display the phosphorylation status of cardiac troponin I determined by phos-tag gel analysis before and after incubation with PKA. 0P <t>represents</t> <t>non-phosphorylated</t> cTnI, 1P single-phosphorylated cTnI and 2P double-phosphorylated cTnI. (G) shows the phos-tag gel image for the quantified data in (H). (I-K) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 compared to donor. (L-N) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 before and after PKA treatment. * p < 0.05, ** p < 0.01, unpaired t -test. n(Donor) = 4, n(HCM 173) = 4/3, n(HCM 175) = 4/3, R278C = 3/3 for with/without PKA, respectively.
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(A-C) show myofilament Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation compared to donor tissue. (D-F) show Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation before and after normalizing <t>cTnI</t> phosphorylation by incubation with protein kinase A (PKA). (G) and (H) display the phosphorylation status of cardiac troponin I determined by phos-tag gel analysis before and after incubation with PKA. 0P <t>represents</t> <t>non-phosphorylated</t> cTnI, 1P single-phosphorylated cTnI and 2P double-phosphorylated cTnI. (G) shows the phos-tag gel image for the quantified data in (H). (I-K) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 compared to donor. (L-N) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 before and after PKA treatment. * p < 0.05, ** p < 0.01, unpaired t -test. n(Donor) = 4, n(HCM 173) = 4/3, n(HCM 175) = 4/3, R278C = 3/3 for with/without PKA, respectively.
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(A-C) show myofilament Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation compared to donor tissue. (D-F) show Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation before and after normalizing <t>cTnI</t> phosphorylation by incubation with protein kinase A (PKA). (G) and (H) display the phosphorylation status of cardiac troponin I determined by phos-tag gel analysis before and after incubation with PKA. 0P <t>represents</t> <t>non-phosphorylated</t> cTnI, 1P single-phosphorylated cTnI and 2P double-phosphorylated cTnI. (G) shows the phos-tag gel image for the quantified data in (H). (I-K) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 compared to donor. (L-N) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 before and after PKA treatment. * p < 0.05, ** p < 0.01, unpaired t -test. n(Donor) = 4, n(HCM 173) = 4/3, n(HCM 175) = 4/3, R278C = 3/3 for with/without PKA, respectively.
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(A-C) show myofilament Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation compared to donor tissue. (D-F) show Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation before and after normalizing <t>cTnI</t> phosphorylation by incubation with protein kinase A (PKA). (G) and (H) display the phosphorylation status of cardiac troponin I determined by phos-tag gel analysis before and after incubation with PKA. 0P <t>represents</t> <t>non-phosphorylated</t> cTnI, 1P single-phosphorylated cTnI and 2P double-phosphorylated cTnI. (G) shows the phos-tag gel image for the quantified data in (H). (I-K) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 compared to donor. (L-N) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 before and after PKA treatment. * p < 0.05, ** p < 0.01, unpaired t -test. n(Donor) = 4, n(HCM 173) = 4/3, n(HCM 175) = 4/3, R278C = 3/3 for with/without PKA, respectively.
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(A-C) show myofilament Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation compared to donor tissue. (D-F) show Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation before and after normalizing <t>cTnI</t> phosphorylation by incubation with protein kinase A (PKA). (G) and (H) display the phosphorylation status of cardiac troponin I determined by phos-tag gel analysis before and after incubation with PKA. 0P <t>represents</t> <t>non-phosphorylated</t> cTnI, 1P single-phosphorylated cTnI and 2P double-phosphorylated cTnI. (G) shows the phos-tag gel image for the quantified data in (H). (I-K) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 compared to donor. (L-N) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 before and after PKA treatment. * p < 0.05, ** p < 0.01, unpaired t -test. n(Donor) = 4, n(HCM 173) = 4/3, n(HCM 175) = 4/3, R278C = 3/3 for with/without PKA, respectively.
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Image Search Results


(A-C) show myofilament Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation compared to donor tissue. (D-F) show Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation before and after normalizing cTnI phosphorylation by incubation with protein kinase A (PKA). (G) and (H) display the phosphorylation status of cardiac troponin I determined by phos-tag gel analysis before and after incubation with PKA. 0P represents non-phosphorylated cTnI, 1P single-phosphorylated cTnI and 2P double-phosphorylated cTnI. (G) shows the phos-tag gel image for the quantified data in (H). (I-K) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 compared to donor. (L-N) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 before and after PKA treatment. * p < 0.05, ** p < 0.01, unpaired t -test. n(Donor) = 4, n(HCM 173) = 4/3, n(HCM 175) = 4/3, R278C = 3/3 for with/without PKA, respectively.

Journal: Journal of molecular and cellular cardiology

Article Title: Mutation location of HCM-causing troponin T mutations defines the degree of myofilament dysfunction in human cardiomyocytes

doi: 10.1016/j.yjmcc.2020.10.006

Figure Lengend Snippet: (A-C) show myofilament Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation compared to donor tissue. (D-F) show Ca 2+ -sensitivity measured as EC 50 for the human samples HCM 173, HCM 175 and HCM 234 carrying the TNNT2 R278C mutation before and after normalizing cTnI phosphorylation by incubation with protein kinase A (PKA). (G) and (H) display the phosphorylation status of cardiac troponin I determined by phos-tag gel analysis before and after incubation with PKA. 0P represents non-phosphorylated cTnI, 1P single-phosphorylated cTnI and 2P double-phosphorylated cTnI. (G) shows the phos-tag gel image for the quantified data in (H). (I-K) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 compared to donor. (L-N) Length-dependent activation of the human samples HCM 173, HCM 175 and HCM 234 before and after PKA treatment. * p < 0.05, ** p < 0.01, unpaired t -test. n(Donor) = 4, n(HCM 173) = 4/3, n(HCM 175) = 4/3, R278C = 3/3 for with/without PKA, respectively.

Article Snippet: Non-, mono- and bis-phosphorylated forms of cTnI (Pierce, MA1-22700) were quantified by phos-tag gel analysis as described previously [ ].

Techniques: Mutagenesis, Phospho-proteomics, Incubation, Activation Assay