Fig. 6

In-vivo H19 inhibition alters HDAC6 and IRS1 levels in the skeletal muscle of mice. A Mice were injected (i.v) with either the scramble or H19 siRNA (3 mg/kg body weight) as described in a previous study. Skeletal muscle tissues were isolated and the levels of H19 were analyzed by qRT-PCR. 18S rRNA was taken as the normalization control. B Skeletal muscle tissues from scramble and H19 siRNA injected mice were lysed and 90 µg of tissue was assayed for HDAC activity as described in the “Materials and methods” section. Values obtained were normalized to the total protein content. Total RNA (1 µg) isolated from skeletal muscle tissues of both groups of mice was reverse transcribed and the transcript levels of Hdac6 (C) and Irs1 (D) were assessed by qRT-PCR. 18S rRNA was taken as the loading control. Mice were fed either chow diet or high fat diet and after 6 months, skeletal muscle tissues were isolated total RNA extracted (1 µg), was reverse transcribed and the transcript levels of H19 (E), Hdac6 (F) and Irs1 (G) were evaluated by qRT-PCR. 18S rRNA was used as the loading control. Values presented are means ± SD of at least four animals in each group. *p ≤ 0.05; **p ≤ 0.01