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. 2018 May 1;84(10):e00235-18. doi: 10.1128/AEM.00235-18

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FIG 4 — TLC and mass spectrometry of 12α-hydroxysteroid dehydrogenase overnight reaction products. In lanes 1 and 2, DCA and 12-oxoLCA were spotted as standards, respectively. Purified recombinant protein (12.5 nM rCHYL, 8 nM rCSCI, 8 nM rCHIR) was incubated overnight under the following conditions: lane 3, 12-oxoLCA plus NADPH, no enzyme; lane 4, 12-oxoLCA plus NAD⁺ plus enzyme; lane 5, 12-oxoLCA plus NADP⁺ plus enzyme; lane 6, 12-oxoLCA plus NADH plus enzyme; lane 7, 12-oxoLCA plus NADPH plus enzyme. Arrow depicts spot analyzed by electrospray ionization mass spectrometry in negative mode, and compared to authentic DCA standard. Formula weight for DCA is 392.58. (a to c) TLC of CSCI (a), CHYL (b), and CHIR (c) reaction products.