Journal: Scientific Reports
Article Title: Defective regulation of POMC precedes hypothalamic inflammation in diet-induced obesity
doi: 10.1038/srep29290
Figure Lengend Snippet: NPY/POMC ( A,B ) and IL6/TNF α ( C,D ) transcripts levels were measured in the hypothalamus of obese-prone (OP) and obese-resistant (OR) mice fed on a high-fat diet for 12 and 24 h, respectively, and samples were collected without fasting. Panel E depicts the real-time PCR determination of NPY and POMC transcript expression in the hypothalamus of mice treated either with NPY or POMC small-interfering RNAs (siRNA); control expression was obtained by the treatment with scramble siRNA. Panel F depicts the protocol employed for experiments presented in panels G–N. For that, OP mice were treated either with a scramble or an anti-NPY small interfering RNA (sc or siNPY, respectively) and OR mice were treated either with a scramble or an anti-POMC small interfering RNA (sc or siPOMC, respectively); food intake was determined before ( G ) and 24 h ( H ) three days ( I ) and seven days ( J ) after siRNA treatment; NPY/POMC transcripts were determined in the hypothalamus three ( K ) or seven ( L ) days after siRNA treatment; TNF α ( M ) and IL6 ( N ) transcripts levels were measured in the hypothalamus of OR mice treated either with a scramble or an anti-POMC small interfering RNA (sc or siPOMC, respectively). In ( O,P ) β-endorphin and α -MSH, respectively, were determined using ELISA in hypothalamic protein extracts obtained 12 or 24 h after introduction of a high-fat diet for OP and OR mice. In ( Q ) the variations of rat blood β-endorphin levels, determined using ELISA, from base-line to 12 h after introduction of a high-fat diet were plotted against body mass gain during a four-week period feeding on a high-fat diet. In ( A–D ) the results are presented as relative to the level of respective transcripts expressed in the hypothalamus of mice fed on chow (dashed line). In E, the results are presented as transcript expression relative to scramble treated controls. In ( O,P ) dashed line indicates values of mice fed on chow. In all experiments n = 6; in ( A–D ) *p < 0.05 vs. chow; § p < 0.05 vs. respective OR; in ( E ) *p < 0.05 vs. respective scramble treated controls; in ( G–N ) *p < 0.05 vs. respective sc; in ( O–P ) *p < 0.05 vs. chow; § p < 0.05 vs. respective 12 h.
Article Snippet: NPY siRNA (sc-42100), POMC siRNA (sc-37278) and control siRNA (sc-37007) were from Santa Cruz Biotechnology, Inc.
Techniques: Real-time Polymerase Chain Reaction, Expressing, Control, Small Interfering RNA, Enzyme-linked Immunosorbent Assay
