Analysis of Bcl-2 family members and apoptosis in murine hematopoietic progenitor cell lines after growth factor withdrawal.
(A) Western blot analysis of Hrk, Bcl-xL, Bax, Bad, and Bak in cells cultured in the presence (0 hours) or absence of IL-3 (FDCP-Mix, FL5.12) or erythropoietin (HCD-57) for 24 and 48 hours. (B) Semiquantitative RT-PCR analysis of Hrk and Bcl-xL mRNA at 0, 24, and 48 hours after growth factor deprivation. PCR products were electrophoresed onto a 2% agarose gel. β-actin mRNA was used as an amplification control. (C) DNA fragmentation analysis in cells cultured with or without growth factor. Cells were incubated for the indicated time points and genomic DNA fragmentation was monitored by electrophoresis onto a 2% agarose gel and staining with ethidium bromide.
