Journal: Nature medicine

Article Title: Targeting the MLL complex in castration resistant prostate cancer

doi: 10.1038/nm.3830

Figure Lengend Snippet: Androgen receptor interacts with MLL complex proteins. ( a ) VCaP nuclear lysate fractions from Superose-6 gel filtration column were immunoblotted using indicated antibodies. Arrows indicate the approximate mass of the complexes eluted. Distribution of cytoplasmic or nuclear proteins demonstrates the efficiency of lysate fractionation (inset western blot). Shown are representative blots ( n =3). ( b–c ) VCaP ( b ) and LNCaP ( c ) nuclear lysates were immunoprecipitated (IP) with AR, ASH2L and menin antibodies followed by immunoblotting (IB) using indicated antisera. Shown are representative blots from three independent experiments. ( d ) VCaP cells were fixed and immunostained using AR (green) and ASH2L (red) or menin (red) antibodies. Confocal microscopic images were analyzed by ImageJ software. Co-localization is represented by yellow puncta. Bar = 10μm. Inset bar = 2.5μm. Shown are representative images from three independent experiments.

Article Snippet: 5mg of nuclear protein was concentrated to 500ul using Microcon centrifugal filter (Millipore) and then applied to a Superose 6 size exclusion column (10/300 GL GE Healthcare) pre-calibrated using the Gel Filtration HMW Calibration Kit (GE Healthcare).

Techniques: Filtration, Fractionation, Western Blot, Immunoprecipitation, Software