Nutritional Regulation of Dies1 and Lipolysis Assay.

<p><b><i>A. Analysis of Dies1 Transcript in WAT with Obesity and Nutritional Status.</i></b> A. qPCR analysis of Dies1 transcript level in murine WAT in <i>ob/ob</i> genetic obesity model (left panel), high fat diet (HFD) <i>vs.</i> lean diet (Reg) (middle panel) and in fasted (Fast) and refed murine (RF) WAT (right panel). Value of the first column in each respective graph was set to 1. For WT n = 2; for <i>ob/ob</i> n = 2; for Reg n = 3; for HFD, n = 3; for fasted n = 2, and for refed n = 2. <b><i>B. Effect of Dies1 Knockdown on Lipolysis.</i></b> 3T3-L1 adipocytes transfected with the indicated siRNAs were assessed for glycerol release into culture media. Iso indicates isoproterenol treated. For <b><i>A</i></b>, * indicates <i>p</i><0.05 compared to the first column of respective graph and N.S. indicates <i>p</i>>0.05. For <b><i>B</i></b>, * indicates <i>p</i><0.05 compared to first column (siCon) and ** indicates <i>p</i><0.05 compared to second column (siCon-iso). For <b>7</b><b><i>B</i></b>, transfections of indicated siRNAs were conducted in at least quadruplicate wells and glycerol assay done in duplicate <i>per</i> sample well. A.U., arbitrary units. The overall experiment was conducted twice with essentially the same results; one such study is presented.