NextSeq 500 paired end sequencing; Thr172 phospho-AMPK ChIP-seq in mouse liver
Thr172 phospho-AMPK ChIP-seq in mouse liver
2 mice for each condition. input is a pool of 2 input samples from each condition. 5 µg of rabbit anti-p-AMPK (Thr 172) ) (Santa Cruz 33524) or normal rabbit IgG antibodies used for ChIP. C57BL/6J male mice of 8 week-old were obtained from Charles River.  In the experiment 1, 4 mice male were fasted 16 h, after which either placebo or a combination of water-soluble DEX (2 mg/kg) and GW (2 mg/kg) was given i.p. Four hours later, mice were sacrificed and the liver was collected for ChIP. In the experiment 2, 4 male mice underwent a 16 h starvation regimen. Then, 2 mice allowed a refeeding (fed group) whilst the other 2 mice stayed fasting (fast group). 1 hour later, mice were sacrificed and livers were collected for ChIP. DNA was purified with a QIAquick PCR purification kit (Qiagen, 28106) according to the user's manual.  NEBNext Ultra DNA Libr Prep. VIB Nucleomics Core facility
Experimental Factor: stimulus
overnight starvation + 1 h refed
Experimental Factor: immunoprecipitate
anti-p-AMPK
Experimental Factor: compound
none