Figure 4
Gene expression in WT and Jdp2KO MEFs as determined by western blotting, microarray analysis and real-time PCR. (a) Western blots of cell-cycle-related proteins in MEFs from WT and Jdp2KO mice. MEFs were synchronized by serum starvation for 48 h and then re-stimulated with 15% FCS. MEFs were harvested 8, 12 and 16 h after addition of serum and cell lysates were prepared. (b) Summary of the microarray data obtained with mRNA from WT and Jdp2KO MEFs. (c) Validation of hybridization intensities. The correction coefficient with respect to relative changes in mRNA levels in two assays was higher than 0.8, indicating that the microarray data are valid. (d) Analysis by quantitative PCR of the expression of representative genes using mRNA from WT MEFs (gray lanes) and Jdp2KO MEFs (black lanes). MEFs were harvested after serum induction for 3 and 18 h, and then real-time PCR was performed with primer sets for the indicated genes, as described in the text. The level of each respective mRNA in unstimulated WT MEF was taken as 100. FCS, fetal calf serum; KO, Jdp2-knockout; MEF, murine embryonic fibroblast; WT, wild type.
