Analysis of mRNA expression of BH3-only genes.

To profile mRNA expression of BH3-only genes by semiquantitative RT-PCR, granulocytes were derived from (A) bone marrow or (B) the peritoneal cavity of C57BL/6 mice, sorted by flow cytometry, and cultured in the presence or absence of rh G-CSF (100 U/mL). RNA was extracted at the indicated time points and reverse transcribed into cDNA. Limiting dilutions of cDNAs were amplified by PCR. The PCR products obtained using BH3-only gene sequence–specific primers were separated on 2% agarose gels and the identity of the PCR products confirmed by Southern blotting using sequence-specific internal oligonucleotides as a probe. Results shown are representative of 2 independent experiments.