Figure 2. SBM does not inhibit mitotic clonal expansion.

(A) Differentiation of 3T3-L1 cells was induced and cell number was counted in control and SBM-treated cells using a hemacytometer 24 h after induction. (B) Effect of SBM on DNA synthesis was monitored by [³H]thymidine incorporation after induction of differentiation of 3T3-L1 in the presence of [³H]thymidine for 24 h. Incorporation of [³H]thymidine into newly synthesized DNA was quantitated with a scintillation counter. (C) mRNA expression of adipogenic transcription factors C/EBPα, C/EBPβ, C/EBPδ, PPARγ, CD36, LPL, aP2 and FAS after 1, 4, and 8 days of differentiation in control and SBM-treated cells. Results were expressed relative to untreated cells after normalization to 18S rRNA. Total cell mass was assessed for different parameters. Data are expressed as the mean ± SD. *P<0.01, **P<0.05 vs. controls. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. Black bars  =  −SBM, gray bars  =  +SBM.