Lack of effect of nonspecific differentiation upon demethylation. (A) Surface expression of GPVI by UT-7/TPO cells cultured in the presence of GM-CSF. UT-7/TPO cells were cultured in the presence of 1 ng/mL GM-CSF for 2 days, and surface expression of GPVI was analyzed by flow cytometry using antibody 204-11 (—). --- corresponds to staining with fluorescein isothiocyanate-labeled secondary antibody alone. The respective GMFI for each tracing is indicated in the respective panel, and these results are representative of 3 independent experiments. (B) Methylation-specific PCR of the GP6 promoter in UT-7/TPO cells cultured in the presence of GM-CSF. The reverse primer msp-R2 was used to specifically amplify methylated alleles. A product was obtained when UT-7/TPO were cultured in the presence of GM-CSF (right lane) but not when they were cultured in the presence of TPO (center lane).