(A) PU.1 mRNA expression in parental and retrovirus-infected 32Dcl3 cells. Total RNA was isolated from infected 32D/LXSN cells (lane 2) or 32D PU.1 cells (lane 3) after selection in G418, and 5 μg from each sample was electrophoresed, blotted, and hybridized to a 32P-labeled PU.1 cDNA probe. Parental 32D cells (lane 1) were used as control. Arrows indicate the position of endogenous and exogenous PU.1 mRNAs. (B) Western blot analysis of PU.1 expression in 32D/PU.1 cells. Total lysates of parental 32Dcl3 cells (lane 1), and cells infected with the LXSN retrovirus (32D/LXSN; lane 2) or with the LXSN retrovirus carrying the PU.1 cDNA (32D/PU.1, lane 3) were subjected to SDS-PAGE, transferred onto nitrocellulose, and incubated with an anti-PU.1 polyclonal antibody. The membrane was then stripped and incubated with an anti–β-actin antibody as a control for equal loading.
