Measurement of the H2S production capacity in tissues was performed as previously described.17 (link),27 (link) In brief, 80 μg of protein was incubated in 10 mM L-cysteine and 1 mM pyridoxal 5′-phosphate hydrate (Sigma-Aldrich, St Louis, MO), or 20 μL of plasma was incubated in 100 mM L-cysteine and 10 mM pyridoxal 5′-phosphate hydrate. This mixture was sealed under lead acetate paper and incubated until black lead sulfide precipitate was detected, but not saturated. The lead sulfide presence was quantified using Fiji software, version 1.53f51 (available at: http://fiji.sc/Fiji ).
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