Phenylhydrazine hydrochloride phz

Haemolytic anaemia was induced in adult GULO^-/- mice (Figure 5A) maintained on either high (1g/L) or low (0.1g/L) vitamin C supplementation. Anemia was induced by phenylhydrazine hydrochloride (PHZ, Sigma-Aldrich) diluted in PBS. Mice were injected intraperitoneally at days 0, 1, and 3, with doses of 40mg/kg, 20mg/kg, and 20mg/kg, respectively. Haematocrits were measured in retroorbital samples on days 2 and 5. Mice were euthanized on day 5 for analyses.

RPMI-1640 medium (product no. 30-2001), fetal bovine serum (product no. 30-2020), dimethylsulfoxide (product no. 4-X), and penicillin/streptomycin (100 U/mL) (product no. 30-2300) were purchased from the American Type Culture Collection (ATCC) (Manassas, VA, USA). Hemopexin (Hx) (heme scavenger; product no. 16-16-080513) was obtained from Athens Research and Technology (Athens, GA). Hemin (ferric chloride heme; product no. H9039), phenylhydrazine hydrochloride (PHZ) (for induction of hemolytic anemia; product no. 114715), phorbol-12-myristate-13 acetate (PMA) (product no. P8139), Tak-242 (product no. 614316), IL-13 (product no. SRP3274), LPS (product no. L4391), IFN-ϒ (product no. SRP3058), IL-4 (product no. SRP3093), DNase I (product no. 10104159001), Collagenase D (product no. 11088866001), and Dispase II (product no. D4693) were obtained from Sigma-Aldrich (St. Louis, MO, USA). RBC Lysis Buffer (10×) (product no. 420302) and Cell Staining Buffer (product no. 420201) were obtained from BioLegend (San Diego, CA, USA).