Figure 2. FAM83F and CK1α protein abundance is reduced at the plasma membrane upon IMiD treatment.

(A) Wide-field immunofluorescence microscopy of HCT116 ^GFP/GFPFAM83F knock-in cells, treated with or without 10 μM pomalidomide for 24 h, stained with antibodies recognising GFP (far left panels, green), and CK1α (second row of panels from left, magenta) as well as DAPI (third row of panels from left, blue). Overlay of GFP, CK1α, and DAPI images as a merged image is shown on the right. Immunofluorescence images were captured with a 60× objective. Scale bar represents 10 μm. Two representative images for each staining are shown. (B) Specific subcellular fractions from cytoplasmic, nuclear, and membrane compartments from DLD-1 wild-type and FAM83F^−/− cells treated with or without 10 μM pomalidomide for 24 h, were resolved by SDS–PAGE and subjected to Western blotting with the indicated antibodies. Specificity of cytoplasmic, nuclear and membrane fractions were determined with Western blotting with compartment specific antibodies: α-tubulin (cytoplasmic), Lamin A/C (nuclear), and Na/K-ATPase (membrane).

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