3T3-L1 Preadipocytes were maintained in growth medium (Undifferentiated), or induced for adipocyte differentiation in the absence (Differentiated control) or presence of various concentrations of 1 or rosiglitazone.
CTGF levels (Fig. 5A) in the differentiated control show a significant increase relative to undifferentiated control at day 3 and day 6 (13.2 ±.3% and 41.7 ± 5% respectively; p ≤ 0.001, n = 6).
However, 804 genes were differentially expressed between terminally differentiated control and Dali knockdown cells, of which 376 genes (46.8%) also differed in expression between Dali knockdown and control cells prior to their differentiation.
During the experiments, undifferentiated cells were used as an untreated control (undifferentiated control) and MDI-induced cells (differentiated control, 0.1% DMSO) were used as a vehicle control of basal differentiation.
Although differentiated control cells demonstrate a significant degree of nuclear translocation of unphosphorylated β-catenin, HPRT-knockdown cells reproducibly are markedly deficient in nuclear translocation.
Differentiated control cells showed a dense microtubule array oriented parallel to the longitudinal axis of the cell.
Interestingly, expression of IGF-1 was upregulated also in differentiated control cells, indicating that DMSO-induced cardiac differentiation activates endogenous IGF-1 transcription.
A similar pattern was observed at 15 days, albeit with a lower overall level of ALP induction in the differentiated control (74 ± 18.9% relative to untreated control, p ≤ 0.001, n = 6).
ALP showed maximum induction in the differentiated control cells at 9 days, with an increase of 167 ± 32% (p ≤ 0.001) relative to undifferentiated control.
Based on congregated data of two experiments, a significant increase in adipocytes was found compared to the differentiated control at the lowest level tested, namely 3 nM (17.2% for differentiated control vs 22.4% and 22.1% for BDE-47 and TROG, respectively).
