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. 2019 Mar 6;151(6):758–770. doi: 10.1085/jgp.201812208

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© 2019 Keceli et al.

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Figure 7. — PLN cysteine residues 41 and 46 are important for the functional uncoupling of PLN from SERCA2a by HNO. High Five insect cell microsomes containing SERCA2a coexpressed with PLN single cysteine constructs (C41,46A-PLN [single Cys at position at 36; left], C36,46A-PLN [single Cys at position at 41; middle], C36,41A-PLN [single Cys at position at 46; right]) were suspended (0.2 mg total protein/ml) in 250 mM sucrose and 10 mM imidazole, pH 7.0, treated with either vehicle (squares), anti-PLN monoclonal antibody 2D12 (circles), or 100 µM AS (triangles) and incubated at room temperature for 10 min, after which the treated microsomes were assayed for [Ca²⁺]-dependent ATPase activity at 37°C. The data are shown normalized to their respective maximas to better illustrate the AS/HNO-dependent shift in the [Ca²⁺]-dependent activity curve. Symbols are the average of three experiments, and the error bars represent the SD.