Dialysed serum

Rotenone, AICAR (5-aminoimidazole-4-carboxamide riboside), A-769662, metformin, ZLN024 and PT1 were purchased from Tocris and 2-deoxy-D-glucose (2DG) was purchased from Abcam. Oligomycin and antibodies to MLC, pMLC, pHSP27, pAMPKα, pACC were purchased from Cell Signaling Technology. Salicylate was purchased from Sigma. The anti-VE-cadherin and ROKα antibodies were from BD-Biosciences and the anti-RhoA antibody was from Merck Millipore. Glucose-free DMEM and dialysed serum were purchased from Gibco Life Technologies (Paisley, United Kingdom).

MCF-7, T47D, BT-474, ZR751 and HEK293 cells were maintained in RPMI-1640 media (Sigma) containing 10% foetal-calf serum (FCS) (Gibco) and 1% penicillin/streptomycin (Sigma). For estrogen stimulation assays, cells were grown in phenol red-free RPMI-1640 media (Gibco) supplemented with 10% serum stripped FCS (Hyclone) and 1% penicillin/streptomycin for 24 (KDM3A knockdown chromatin immunoprecipitation (ChIP) experiments) or 48 h (ChIP/gene expression/microarray experiments) prior to the addition of 10 nM 17-β-estradiol (E₂) (Sigma) for 45 min (ChIP) or 4 h (gene expression/microarray analysis). MMU2 cells were maintained in phenol red-free RPMI-1640 media supplemented with 10% dialysed serum (Gibco) and 1% penicillin/streptomycin. MCF-10A cells were maintained in DMEM-F12 (Sigma) media containing 5% horse serum (Sigma), 10 μg/ml insulin (Sigma), 0.5 μg/ml EGF (Sigma), 100 ng/ml cholera toxin (Sigma) and 1% penicillin/streptomycin.