Rabbit control igg

Sodium palmitate (#P9767) and Oil Red O (ORO, #O0625) were purchased from Sigma-Aldrich (Milwaukee, WI, USA). Dimethyl sulfoxide (DMSO, #D8371), fatty acid free bovine serum albumin (BSA, #A8850) and Nile red (#N8440) was obtained from Solarbio (Beijing, China). Radicicol (#HY-N6769) was purchased from MedChemExpress (Monmouth Junction, NJ, USA). The primary antibodies against LKB1(3047s, 1:1000), phospho-LKB1^Ser428 (3482s, 1:1000), AMPKα (2732s, 1:1000), phospho-AMPKα^Thr172 (2535s, 1:1000), ACC (3676s, 1:1000), phospho-ACC^Ser79 (11818s, 1:1000), FASN (3180s, 1:1000) and β-actin (4970s, 1:1000) were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies for CPT1α (ab128568, 1:1000), MO25 (ab51132, 1:10000) and STRAD (ab192879, 1:2000) were bought from Abcam (Cambridge, MA, USA). Antibodies against LKB1(10746-1-AP) were obtained from Proteintech (Rosemont, IL, USA). Anti-SREBP1 (sc-13551, 1:200) and anti-Nitro-tyrosine (sc-32757) were purchased from Santa Cruz Biotechnology (Santa Cruz, Dallas, Texas, USA). Alexa Fluor^® 488 conjugated WGA (#W11261) was obtained from Thermo Fisher Scientific (Waltham, MA, USA). Rabbit Control IgG (#AC005) and Mouse Anti-Rabbit IgG Light Chain (#AS061, 1:5000) were purchased from ABclonal Biotechnology (Wuhan, China).

The following three antibodies were used for ChIP experiments: H3K27ac (Abcam, #AB4729), E2F7 Polyclonal Antibody (ThermoFisher Scientific, #A303-037A-T) and Rabbit Control IgG (Abclonal, #AC005). ChIP assay was performed with 2 × 10⁷ adherent cells lysed to prepare nuclear extracts. Firstly, cells were treated with 1% formaldehyde to crosslink DNA. After chromatin shearing by sonication, the nuclear lysates were treated at 4°C overnight with protein A Dynabeads (Invitrogen, USA) combined with 3-5 g of antibody to prepare each sample. The beads were then retrieved with a magnet and cleaned. The DNA was then decrosslinked for 4 hours at 55°C, and purified by QIAquick PCR Purification Kit (QIAGEN, USA). 5-10 ng of pure ChIP DNA was utilized as input material for subsequent detection in each sample. At last, DNA collected from the experiments was examined by qRT-PCR assays.