Fig. 5

Effects of PPE on the expressions of C/EBPβ and C/EBPδ, and IDM-induced mitotic clonal expansion. a 3T3-L1 cells which reached confluence were cultured with fraction No. 9 and IDM for 12 h. The time-course (0, 0.5, 1, 2, 4, 6 and 12 h) of the gene expressions of C/EBPβ and C/EBPδ was analyzed by RT-qPCR. The gene expressions are presented relative to the value for 3T3-L1. Experiments were performed in triplicate, and the data are presented as the mean ± SEM (n = 3). b 3T3-L1 cells which reached confluence were cultured with fraction No. 9 and IDM for 48 h. The time-course (0, 1, 3, 6, 12, 24 and 48 h) of the protein expressions of C/EBPβ and C/EBPδ was analyzed by western blotting. Vehicle represents cells cultured with PBS and IDM, and 3T3-L1 represents cells cultured without both fraction No. 9 and IDM. Experiments were repeated three times, and representative results are shown. c 3T3-L1 cells which reached confluence were cultured with IDM in the presence of fraction No. 9 (IDM + Fraction No. 9) or PBS (IDM + Vehicle) for 24 h. 3T3-L1 represents cells cultured without either fraction No. 9 or IDM for 24 h. The cells were stained with propidium iodide and then analyzed with flow cytometry. The experiments were repeated three times, and representative results are shown. d Bar diagram depicting the percentage of cells in different phases (G0/G1, S, G2/M) of the cell cycle. e 3T3-L1 cells which reached confluence were cultured for 48 h with IDM and a two-fold dilution series of fraction No. 9. Control represents cells cultured with only IDM, and 3T3-L1 represents cells cultured without either PPE or IDM. The cell proliferation data are presented as a percentage of the value in Control cells. Experiments were performed in triplicate, and the data are presented as the mean ± SEM (n = 3). *p < 0.05, **p < 0.01 vs. Control. Experiments were repeated at least three times, and representative results are shown