Increased amount of antiapoptotic and cell cycle–related proteins in purified wt and thalassemic erythroid cells. Representative Western blots performed on cells from wt (lanes 1,2), th3/+ (lanes 3,4), and th3/th3 (lanes 5,6) mice, and control cell lines (lane 7) probed with (A) Bcl-XL (control: NIH-3T3 cells), (B) CycA (control: Mel cells); (C) Cdk2 (control: Mel cells); and (D) EpoR (control: K562 cells). The upper band in panel A is described as the deamidated form of the protein. Bcl-XL deamidation has been shown to produce a complete loss of the antiapoptotic function of Bcl-XL.48 Similar ratios of the 2 bands are present in both normal and thalassemic mice. The membrane used for Bcl-XL was reprobed with CycA antibody. Specific antibodies against the phosphorylated and nonphosphorylated forms of the protein were used sequentially. In all cases, the same membranes were reprobed against β-actin as a loading control.
