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Recent studies have demonstrated that H<jats:sub>2<\/jats:sub>S promotes aerobic energy production in the mitochondria in response to hypoxia, but its effect on anaerobic energy production has yet to be established. Glycolysis is the anaerobic process by which ATP is produced through the metabolism of glucose. Mammalian red blood cells (RBCs) extrude mitochondria and nucleus during erythropoiesis. These cells would serve as a unique model to observe the effect of H<jats:sub>2<\/jats:sub>S on glycolysis\u2010mediated energy production. The purpose of this study was to determine the effect of H<jats:sub>2<\/jats:sub>S on glycolysis\u2010mediated energy production in mitochondria\u2010free mouse RBCs. Western blot analysis showed that the only H<jats:sub>2<\/jats:sub>S\u2010generating enzyme expressed in mouse RBCs is 3\u2010mercaptopyruvate sulfurtransferase (MST). Supplement of the substrate for MST stimulated, but the inhibition of the same suppressed, the endogenous production of H<jats:sub>2<\/jats:sub>S. Both exogenously administered H<jats:sub>2<\/jats:sub>S salt and MST\u2010derived endogenous H<jats:sub>2<\/jats:sub>S stimulated glycolysis\u2010mediated ATP production. The effect of NaHS on ATP levels was not affected by oxygenation status. On the contrary, hypoxia increased intracellular H<jats:sub>2<\/jats:sub>S levels and MST activity in mouse RBCs. The mitochondria\u2010targeted H<jats:sub>2<\/jats:sub>S donor, AP39, did not affect ATP levels of mouse RBCs. NaHS at low concentrations (3\u2013100\u2009\u03bcM) increased ATP levels and decreased cell viability after 3 days of incubation in vitro. Higher NaHS concentrations (300\u20131000\u2009\u03bcM) lowered ATP levels, but prolonged cell viability. H<jats:sub>2<\/jats:sub>S may offer a cytoprotective effect in mammalian RBCs to maintain oxygen\u2010independent energy production.<\/jats:p>","DOI":"10.1002\/jcp.30544","type":"journal-article","created":{"date-parts":[[2021,8,4]],"date-time":"2021-08-04T07:31:11Z","timestamp":1628062271000},"page":"763-773","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Effect of hydrogen sulfide on glycolysis\u2010based energy production in mouse erythrocytes"],"prefix":"10.1002","volume":"237","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1841-3482","authenticated-orcid":false,"given":"Eden T.","family":"Wondimu","sequence":"first","affiliation":[{"name":"Cardiovascular and Metabolic Research Unit Laurentian University Sudbury Ontario Canada"},{"name":"Department of Biology Laurentian University Sudbury Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Quanxi","family":"Zhang","sequence":"additional","affiliation":[{"name":"Cardiovascular and Metabolic Research Unit Laurentian University Sudbury Ontario Canada"},{"name":"School of Life Science Shanxi University Taiyuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuping","family":"Jin","sequence":"additional","affiliation":[{"name":"Cardiovascular and Metabolic Research Unit Laurentian University Sudbury Ontario Canada"},{"name":"School of Life Science Shanxi University Taiyuan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ming","family":"Fu","sequence":"additional","affiliation":[{"name":"Cardiovascular and Metabolic Research Unit Laurentian University Sudbury Ontario Canada"},{"name":"School of Human Kinetics Laurentian University Sudbury Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberta","family":"Torregrossa","sequence":"additional","affiliation":[{"name":"University of Exeter Medical School Exeter UK"},{"name":"MitoRx Therapeutics Oxford UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthew","family":"Whiteman","sequence":"additional","affiliation":[{"name":"University of Exeter Medical School Exeter UK"},{"name":"MitoRx Therapeutics Oxford UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3575-7786","authenticated-orcid":false,"given":"Guangdong","family":"Yang","sequence":"additional","affiliation":[{"name":"Cardiovascular and Metabolic Research Unit Laurentian University Sudbury Ontario Canada"},{"name":"Department of Chemistry and Biochemistry Laurentian University Sudbury Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingyun","family":"Wu","sequence":"additional","affiliation":[{"name":"Cardiovascular and Metabolic Research Unit Laurentian University Sudbury Ontario Canada"},{"name":"School of Human Kinetics Laurentian University Sudbury Ontario Canada"},{"name":"Health Sciences North Research Institute Sudbury Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3825-3620","authenticated-orcid":false,"given":"Rui","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Biology York University Toronto Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2021,8,4]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ejphar.2005.12.037"},{"key":"e_1_2_9_3_1","volume-title":"Medical biochemistry","author":"Baynes J. 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