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. 2020 Aug 31;295(45):15342–15365. doi: 10.1074/jbc.RA120.014467

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© 2020 Diering et al.

Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.

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Figure 6. — Identification and validation of proteins enriched by cAMP-agarose pulldown. ARVMs were treated with vehicle (DMSO), NCA (100 µmol/liter, 30 min), CXL-1020 (300 µmol/liter, 15 min), or ISO (10 nmol/liter, 10 min) and harvested under NR or R conditions. A, pulldown experiments using 8-AHA-cAMP-agarose were performed and analyzed by MS. Density chart reveals the number of identified peptides per protein hit. B, Western IB analysis of the pulldown samples for PKA-RI and cMyBP-C was performed under R or NR conditions. The scatter plots represent data of co-precipitated cMyBP-C in the pulldown fraction from 4 independent experiments. Results are expressed as fold-change over the ISO signal. *, p < 0.01 for comparison between the control (vehicle) and the HNO donor-treated group by one-way ANOVA with Dunnett's post-test (F = 8.38, p = 0.0028).