Gain and loss of function assays to validate the role of miR-143 in young (6 mo) and old (30 mo) rat SVF derived cells, post-adipogenesis.

<p>Adipocyte differentiation was performed on the cells after transient transfection of both young and old rat (n = 8) preadipocytes by premir or antagomir-143 (gain or knockdown of miR-143). At the end of 14 days post-adipogenesis, the transfection efficiency and target gene levels were analyzed by RT-qPCR and Western blot analysis. miR-143, <i>8A</i>; ERK5 mRNA, <i>8B</i>; ERK5 protein, <i>8C</i>; PPARg mRNA, <i>8D</i>; Runx2, <i>8E;</i> and miR-204, <i>8F.</i> The real time PCR data was expressed as fold of relative expression±SEM (Standard Error of Mean). *: p<0.05; **: p<0.01; ***: p<0.005.