Figure 2
Proliferative properties of MEFs from WT and Jdp2KO mice. (a) The expression of JDP2 mRNA in embryos and in MEFs from WT and Jdp2KO mice was analyzed by northern blotting. The blot was re-probed for analysis of GPDH mRNA as loading control. (b) The expression of JDP2 in MEFs from WT and Jdp2KO mice was examined by western blotting. (c, d) Proliferation of MEFs from WT and Jdp2KO mice. MEFs were cultured in 10-cm dishes with DMEM plus 10% FCS for 3 days and then re-inoculated at the same density during 20 passages. Mean cell numbers from four independent plates were determined at each passage. A representative assay was shown (c; Trypan blue dye-exclusion test). Similar results were obtained in each experiment of three pairs of independent MEF clones. The number of population doublings (PD) in each passage was calculated as described elsewhere (Polisetty et al., 2008). Number of cell doubling (NCD)=log10(N/N0)/log102, where ‘N’ is the final density of the cells and ‘N0’ is the initial seeding density of the cells. In panel d, MEFs were starved in DMEM plus 0.1% FCS for 36 h and then re-plated in DMEM plus 15% FCS and cultured for 5 days. MEFs were counted (six repeats; Alamar Blue assay). *P<0.05; **P<0.001. (e) MEFs from WT and Jdp2KO mice were plated in gelatin-coated dishes. Colonies (of more than 2 mm in diameter) were counted 2 weeks later. DMEM, Dulbecco's modified Eagle's medium; FCS, fetal calf serum; JDP2, Jun dimerization protein-2; KO, Jdp2-knockout; MEF, murine embryonic fibroblast; WT, wild type.
