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After approximately 16 hours of incubation with hormone, cells in each well were lysed using 100L mammalian protein extraction reagent	After blocking, sections were stained with antibodies or phalloidin and imaged with a Leica DM4000B fluorescent microscope	After centrifugation, the halo containing PBMCs was carefully transferred into a clean centrifuge tube and washed twice with 7 ml of PBS
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After identifying potential binding sites at HA with the MOE module ``Site Finder'', the test compounds were docked into the 30 cavities with the best PLB	After incubation at room temperature for 30 min, liquid of each sample well was removed, washed and subsequently incubated with the color reaction TMB Chromogen in the dark for 15 min at room temperature	After incubation with the appropriate peroxidase-conjugated secondary antibody (DAKO, Denmark), the immunoreactive bands were visualized by chemioluminescence
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