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Material and methods
A rabbit polyclonal antibody to human recombinant AdK was raised in our lab by immunizing New Zealand White rabbits (Charles River). For immunofluorescence labeling, cells growing on coverslips were fixed by immersion in 100% methanol at ?20 °C for 10 min. After blocking nonspecific signals by incubation with 3% BSA in TBS, the cells were incubated in 1:100 dilutions of the primary Torin1 (viz. rabbit polyclonal antibody to AdK or a mouse monoclonal antibody to c-Myc (Cat. No. 2Q329; Santa Cruz Biotechnology) for 2 h at 37 °C. After rinsing the coverslips three times with TBS they were incubated in 1:200 dilutions of secondary antibodies (viz. anti-mouse or anti-rabbit Alexa Fluor 488; Molecular Probes) for 2 h at 37 °C. After further rinsing, the coverslips were mounted on glass slides in 90% glycerol. Photographs were taken on an Olympus BX51 microscope equipped with REITGA Exi (Qimaging) digital camera and Northern Elite software.
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Exposure to A or HIV activates caveolae
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