UCC were treated for 72 h with cell line-specific half maximal inhibitory concentration (IC50) doses of PLX before cell lysates were prepared. Immunoblot analysis was performed with whole cell extracts as described in [8 (link)]. Antibodies for target detection and secondary antibodies are listed in Table S1 . Targets were visualized by SuperSignal™ West Femto (Thermo Scientific, Rockford, IL, USA) and WesternBright Quantum kit (Biozym, Hessisch Oldendorf, Germany).
Immunocytochemistry was performed as described in detail elsewhere [32 (link)]. Briefly, cells were treated on coverslips with the indicated doses, fixed with formaldehyde, permeabilized, and incubated in blocking solution. Cells were stained by the application of 50 μL of primary antibody solution per coverslip (Table S1 ) and incubated at 4 °C overnight. After washing, secondary antibodies were likewise added at room temperature for 1 h. After washing, the nuclei were counterstained with DAPI (4′,6-Diamidin-2-phenylindol). Finally, mounting medium (Dako, Santa Clara, CA, USA) was added. Images were taken by means of an Olympus Fluoview FV-1000 microscope (Hamburg, Germany), kindly provided by the Center for Advanced Imaging, Heinrich Heine University (Cai, numerical aperture 1/20, 60× objective). Images were processed using the Fluoview FV-1000 software, version 3.1.
Immunocytochemistry was performed as described in detail elsewhere [32 (link)]. Briefly, cells were treated on coverslips with the indicated doses, fixed with formaldehyde, permeabilized, and incubated in blocking solution. Cells were stained by the application of 50 μL of primary antibody solution per coverslip (
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