Computational protocol: A possible regulatory link between Twist 1 and PPARγ gene regulation in 3T3-L1 adipocytes

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[…] Western blotting was conducted according to previously described methods []. The membranes were incubated overnight at 4 °C with primary antibodies for anti-PPARγ (1:1,000), anti-Twist1 (1:50) and anti-β-actin (1:2500). The target molecular weights for Twist 1, PPARγ and β-actin were 21 kDa, 58 kDa and 43 kDa, respectively. The relative OD ratio was calculated using ImageJ software and normalized to β-actin. Data from three replicate experiments were analyzed. [...] For Oil Red O staining, the data are presented as the OD values. ImageJ software was used to quantify the results from the western blotting experiment. The relative gray value of each band was calculated based on results of three replicates. All data are shown as the mean ± SD, and the data were analyzed using SPSS software package, version 17.0. T tests were used for comparisons between two groups. A one-way analysis of variance (ANOVA) was performed for comparisons between more than two groups, and Dixon’s q tests were used for further pairwise comparisons. P < 0.05 was considered significant. […]

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