RT-PCR analysis of sex hormone receptors ER , ER β, PR, and AR in the megakaryocytic lineage.

Total RNA was reverse-transcribed and amplified by PCR with the use of primers specific for ER α, ER β, PR, AR, and β actin transcripts. All PCR primer pairs were separated by at least 1 intron to clearly identify products of cDNA amplification. The cells from which RNA was extracted were as follows: CD34⁺ cells prior to treatment with PEG-rhMGDF (D0 [day 0] CD34) (lane 1); CD34⁺ cells treated for 7 days with PEG-rhMGDF (D7 [day 7] CD34) (lane 2); gel-filtered platelets (lane 3); the PC3 prostate cancer cell line (lane 4); the Dami megakaryocytic cell line (lane 5); the HEL megakaryocytic cell line (lane 6); the T47D breast cancer cell line (lane 7). Lanes 8 and 9 contain products from PCRs using genomic DNA or no template, respectively. Lane 10 contains molecular weight marker (Φ × 174 DNA digested with HaeIII endonuclease). Sizes of PCR products are shown in “Materials and methods.”