Bardet-Biedl syndrome protein 4 becomes mislocalized in ciliated cells in response to the CCT suppression. (A) Transfection protocol for transient expression of Δ1–83PhLP-FLAG and phosducin in RPE1-hTERT cells. (B) Representative Western blotting showing the expression of the indicated proteins. (C) RPE1-hTERT cells stably expressing GFP-BBS4 fusion protein were cotransfected with empty- and tdTomato vectors. GFP-BBS4 was immuno-enhanced with antibody against GFP (green), cell nuclei were visualized with DAPI (blue), transfected cells are distinguished from nontransfected cells by the presence of tdTomato fluorescence (red). (D) A number of cells with GFP-BBS4 was determined and expressed as a fraction of all transfected (red) cells. Such fraction in the cells expressing Δ1–83PhLP-FLAG and phosducin was normalized to that found in the control cells treated with empty vector. Error bars are SEM, n = 700, P = 0.008, as determined by paired t-test (***).