Expression of Hrk in FDCP-Mix cells treated with a protein synthesis inhibitor.

(A) IL-3-deprived cells were cultured with or without cycloheximide (CHX), and at the indicated time intervals, the mRNA levels of Hrk were analyzed by semiquantitative RT-PCR. As controls, cells maintained with IL-3 in the absence (first lane) or in the presence (second lane) of CHX were also analyzed and showed no induction of Hrk. PCR products were electrophoresed onto a 2% agarose gel. β-actin mRNA was used as an amplification control. (B) Cells were deprived of IL-3 for 24 hours and then stimulated for different time intervals with IL-3. The expression of Hrk and Bax proteins was determined by Western blotting with use of specific antibodies.