Expression of PRV-1 in granulocytes from healthy volunteers treated with G-CSF in vivo and in vitro.

Granulocytes were isolated from peripheral blood of untreated and G-CSF-treated healthy volunteers and a patient with PV. Total RNA was extracted and a Northern blot prepared using 10 μg RNA. (A) Lane 1 patient with PV, lane 2 empty, lanes 3 and 4 healthy volunteers treated with G-CSF in vivo, and lanes 5 and 6 untreated healthy volunteer donors. (B) Lane 1 unstimulated granulocytes; lanes 2-8, granulocytes stimulated in vitro for the indicated times with 100 ng/mL G-CSF. (Top) The membrane was probed with a 1.1-kb fragment of the PRV-1 cDNA. (Bottom) The membrane was stripped and reprobed with a 1.2 kb fragment of the human β-actin cDNA.