iSLK/RTA-WT, iSLK/RTA-DD, SLK/iBAC.RTA-WT, or SLK/iBAC.RTA-Q37 cells were processed as previously described (53). Briefly, cells were fixed with 4% paraformaldehyde and permeabilized with 1% Triton X-100 buffer. After being blocked with goat serum, cells were incubated with primary mouse monoclonal anti-FLAG M2 antibody or polyclonal anti-RTA (1:100 dilution) serum. Cells were then incubated with Alexa Fluor 488–congugated goat secondary antibody (1:500 dilution), stained with 4′,6-diamidino-2-phenylindole (DAPI), and analyzed with confocal microscope (Nikon). Cells expressing fluorescent fusion proteins were fixed with 4% paraformaldehyde and directly analyzed by immunofluorescence microscope (Nikon). Representative images were shown for all analyses.

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