Figure 6

Ddit4 is induced during fasting and upregulated by p53 activation in cultured adipocytes. (A) In vivo regulation of Ddit4 in fasted and fed mice. Relative expression values from qPCR measurements (n = 5) were normalized to 36b4 expression and the value for the first time point was set to 1. Significance was determined with a 2-way ANOVA followed by a Bonferroni posttest to determine significance for the single time points (*p < 0.05; **p < 0.01; ***p < 0.001). Western blots were performed with tissue samples from six month old mice fed a chow diet (fed) or fasted overnight (fasted). Samples are from three littermates each. The following proteins served as loading controls (L.C.): β-actin for WAT and LIV, β-tubulin for SM. (B) 10 μM Nutlin-3, a specific p53 activator, was used to treat mature C3H10T1/2 adipocytes (day 7 of differentiation) for 6 hours. qPCR mRNA measurements (n = 3) were normalized to 36b4 expression and related to control cells (treated with DMSO). A two-tailed, unpaired student’s t-test was used to determine statistical significance (*p < 0.05; **p < 0.01; ***p < 0.001). The western blot shows data from three independent replicates treated with Nutlin-3 (+) or DMSO (−) using β–actin as loading control.