Figure 3
Up-regulation of Bmf, Noxa, and Puma and down-regulation of Mcl-1 and Bcl-XL by ATO. U266, MM.1s, 8226/S, and KMS11 were treated for 6, 24, and 48 hours with 2 μM ATO. (A) Mitochondrial-rich fractions were obtained and Bmf expression was determined by Western blot. Membranes were reprobed with rabbit anti–COX IV polyclonal antibody to determine loading. (B) Western blot analysis of total protein lysates from ATO-treated cells. Membranes were probed with specific antibodies for Noxa, Puma, Bim, Mcl-1, Bcl-XL, and actin. (C) Cytosolic and heavy membrane fractions were obtained for untreated and 24-hour ATO-treated MM.1s and KMS11 cells as indicated in “Subcellular fractionation.” Actin and COX IV were used as indicators of fraction purity.
