Colocalization analysis of internal D4H signal with components of endo-membranes. Marker proteins were tagged with GFP and expressed together with mCherry-D4H. Localization of both signals was compared in cells treated or not treated with CK-666. Sec24 is a COPII cargo receptor and transitional ER (tER) marker. Anp1 is a subunit of the mannosyltransferase complex that localizes to the early Golgi. Apl5 and Apl6 are subunits of the AP3 adaptor complex present in the TGN network. Vps8 is a subunit of the CORVET (Class C core endosome vacuole tethering) complex present in the prevacuolar compartment. Ypt5 is a Rab GTPase predicted to regulate the CORVET complex. End4 is a clathrin adaptor present at sites of endocytosis. Clc1 encodes the clathrin light chain, which localizes to endosomes/TGN and actin patches. Ypt3 is a Rab GTPase homologous to Rab11, localizing to secretory vesicles and the TGN. Sec3, Sec5, and Exo70 are exocyst components, localizing to secretory vesicles and the PM. Bgs4 is a 1,3-β-glucan synthase present at cell tips and within TGN. Pmp3 is a predicted PM protein, which also labels prominent, unknown internal structures. Bodipy labels lipid droplets. Mitotracker labels mitochondria. No colocalization was observed between any of these markers and D4H internal structures upon CK-666 treatment, except for Clc1, for which partial colocalization was observed. Scale bar, 2 µm.