Western and Northern blot analysis of G-CSF–induced 32Dcl3 cells stably transfected with the pMT2-CDP/cut plasmid. (A) Western blot using anti-CDP antiserum confirming overexpression of CDP/cut in CDP-transfected 32Dcl3 cells before (0) and after (4d) G-CSF induction. 32Dwt18/CDP/cut cells were used as a positive (+) control. Each lane contains 100 μg nuclear protein extracts. (B) Ten micrograms each of total RNA isolated from uninduced or 2-day and 4-day G-CSF–induced 32Dcl3 cells stably cotransfected with pMT2-CDP/cut and pSVneo (lanes 3, 4, and 5, respectively) plasmids or pSVneo plasmid alone (uninduced, lane 1; 4-day G-CSF–induced, lane 2) was electrophoresed on a 1% formaldehyde/agarose gel and blotted onto a nitrocellulose filter. The filter was probed with an [α-³²P]dCTP–labeled LF cDNA probe. The position of 28S and 18S rRNA markers is indicated. LF mRNA is denoted by the arrowhead. To ensure equal loading of RNA in each lane, the blot was stripped and reprobed with an [α-³²P]dCTP–labeled β-actin probe, as indicated in the bottom panel.