![Cysteine maintains GSH levels and the subsequent management of ROS. (A) Relative cell counts at day 2 of HL60 and MOLM13 cells grown with or without cystine, or without cystine but in the presence of GSH. (B) Relative cell counts at day 2 of HL60 and MOLM13 cells grown with or without cystine, or without cystine but in the presence of Tau. (C) Relative cell counts at day 1 of HL60 or MOLM13 cells grown with or without cystine, or without cystine but in the presence of the ROS scavengers: Trolox (100 μM), α-tocopherol (α-toc; 50 μM), D-mannitol (1 mM), Mn(III)tetrakis (4-benzoic acid) porphyrin (MnTBAP, 25 μM), and 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO; 10-100 μM). (D) IC50 curve for cell counts of HL60, MOLM13, and K562 cells grown in a concentration range of the thioredoxin reductase inhibitor auranofin. (E) Relative cell counts at 16 and 24 hours of HL60, MOLM13, and K562 cells grown with or without cystine, or without cystine but in the presence of the NOX inhibitor DPI. (F) Experiment as in panel E, but relative viability is shown. (G) Total cellular ROS (by dichlorofluorescin diacetate [DCFDA]) after 16 and 24 hours of cystine depletion, or after 24 hours cystine depletion but in the presence of DPI. (H) Representative histogram plots for total cellular ROS (DCFDA) of data shown in panel G. Statistical analysis by ordinary 1-way ANOVA; ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001. Rel., relative.](https://ash.silverchair-cdn.com/ash/content_public/journal/bloodadvances/8/1/10.1182_bloodadvances.2023010786/2/blooda_adv-2023-010786-gr3b.jpeg?Expires=1724184088&Signature=fFl1EnInkGThKhGlDPH5FHcjqSOixat5Jv5BUKychukvJa3-l8OhwxrtbG~0XFcNGr5sa4NJvOet0UiB~WJpwkfAQYlionqKyuyTZDyVExIfEE3YAxZ~elINz6v2ms7ByZoRMyj2QNVjJlC0sJsOtNQeagB-pCZBb8tWukzlHlwR~hpSxRD7Zo7LL4OtBD7Z9ZIYI5cAl5P6PKVO1faQZCXIzFIAiQOBNhqe~vYULE5OIGEPqvQo-bob5nD0dat1AEQbFBcWTnGv0ZH3CbvpKHzHn0a6vlocZoL4eb9JLLjBEU9xfdAih~NQS59NW5e1wd0bTl0oFTdaCH8lLUgahQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Cysteine maintains GSH levels and the subsequent management of ROS. (A) Relative cell counts at day 2 of HL60 and MOLM13 cells grown with or without cystine, or without cystine but in the presence of GSH. (B) Relative cell counts at day 2 of HL60 and MOLM13 cells grown with or without cystine, or without cystine but in the presence of Tau. (C) Relative cell counts at day 1 of HL60 or MOLM13 cells grown with or without cystine, or without cystine but in the presence of the ROS scavengers: Trolox (100 μM), α-tocopherol (α-toc; 50 μM), D-mannitol (1 mM), Mn(III)tetrakis (4-benzoic acid) porphyrin (MnTBAP, 25 μM), and 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO; 10-100 μM). (D) IC50 curve for cell counts of HL60, MOLM13, and K562 cells grown in a concentration range of the thioredoxin reductase inhibitor auranofin. (E) Relative cell counts at 16 and 24 hours of HL60, MOLM13, and K562 cells grown with or without cystine, or without cystine but in the presence of the NOX inhibitor DPI. (F) Experiment as in panel E, but relative viability is shown. (G) Total cellular ROS (by dichlorofluorescin diacetate [DCFDA]) after 16 and 24 hours of cystine depletion, or after 24 hours cystine depletion but in the presence of DPI. (H) Representative histogram plots for total cellular ROS (DCFDA) of data shown in panel G. Statistical analysis by ordinary 1-way ANOVA; ∗P < .05, ∗∗P < .01, ∗∗∗P < .001, and ∗∗∗∗P < .0001. Rel., relative.
