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O-dianisidine staining

JP Jin Woo Park
JK Joo-Young Kang
JH Ja Young Hahm
HK Hyun Jeong Kim
SS Sang-Beom Seo
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Hemoglobin activity was detected in K562 cells by performing o-dianisidine staining as described43. Briefly, K562 cells were cultured with 30 nM hemin for 2 days. To detected hemoglobin synthesis, K562 cells were washed with PBS and stained for 15 min in the dark in 0.6 mg/ml 3,3′-dimethoxy benzidine (Sigma, St. Louis, MO, USA), 0.01 M sodium acetate (pH 4.5), 0.65% H2O2 and 40% (v/v) ethanol. The cells were then washed once with PBC and suspended with 70% glycerol in glass slides for microscopy. The percentage of dianisidine-positive cells were calculated by counting three independent fields to obtain the average. For some experiments, K562 cells were co-treated with alisertib (AURKA inhibitor).

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