Figure 5.

Translation of C/EBPα mRNA is impaired in Hzf-knockdown cells. (A) Cells were pulse-labelled with ³⁵S-Met/Cys for the indicated periods, and immunoprecipitated C/EBPα, C/EBPβ, or CDK4 protein was separated by SDS–PAGE and detected by autoradiography. Portions of the total radiolabelled cell extract were analysed in parallel (lower panel). Radioactivity of each protein was measured (right). (B) Cytoplasmic lysates prepared from cells differentiated for 4 days were fractionated using sucrose density gradient centrifugation. rRNA (28S and 18S) was visualized by methylene blue staining. northern blot analysis was performed using C/EBPα or GAPDH cDNA as a probe.