Fig 1: THOC3 is located in cytoplasm and folded by CCT.A Representative images of THOC3 localization in H520/H1703/SK-MES-1 cells detected by Fluorescent In Situ Hybridization (FISH) (40× magnification), scale bars = 100 µm (left); THOC3 mRNA expression in the nucleus and cytoplasm was determined via qRT-PCR in H1703/SK-MES-1/H520 cells (right). B Silver staining of immunoprecipitation of Flag and IgG antibodies in H1703 cells. C Bar chart showing proteins with unused scores of Flag over 10, identified via MS analysis of H1703 cells following immunoprecipitation with Flag and IgG antibodies (top); representative signaling pathways composed of proteins bound to THOC3 based on GO analysis (bottom). D THOC3 expression in cells transfected with si-NC or si-CCT8/CCT6 shown by WB. E Immunoprecipitation of cell lysates from H1703 cells transfected with Flag-tagged THOC3 using anti-CCT8, anti-CCT6A, anti-Flag, or IgG antibody, and immunoprecipitates were blotted with anti-Flag/CCT8/CCT6A antibodies. F Immunofluorescence images showing the distribution of THOC3 (green) and CCT8/CCT6A (red) in H1703 cells. Scale bars = 100 µm. G Cells are transfected with Flag-tagged THOC3 for 18 h and then incubated with and without 50 µg/ml cycloheximide (CHX) for 5 h. Immunoprecipitates were blotted with anti-CCT8/CCT6A antibodies. H Cells expressing Flag tagged THOC3 are prepared in the presence or absence of ATP and MgCl2 and analyzed via immunoprecipitation and WB by anti-Flag/CCT8/CCT6A antibodies.