Localization of GFP-VPS16B within stable megakaryocytic Dami cells. Dami cells stably transfected with GFP-VPS16B fusion protein (green) were costained with different intracellular markers (red) as indicated. Merged images are shown on the right of each row (C,F,I,L,O). Partial colocalization (merge) of GFP-VPS16B is observed with the trans-Golgi network (AP-1, A-C), late endosomes/lysosomes (LAMP1, D-F), late endosomes (Rab7, G-I), α-granules (VWF, J-L) but not δ-granules (CD63, M-O). Insets with arrows highlight areas of colocalization. Multilobed nuclei are readily observed in Dami cells stimulated with phorbol ester and TPO. Representative z-stack images are shown. The white bar represents 5 μm. Images were obtained using a Leica DMIRE2 inverted fluorescence microscope equipped with a Hamamatsu C9100-12 back-thinned EM-CCD camera and Yokogawa CSU 10 spinning disk confocal scan head (with Spectral Aurora Borealis upgrade), with 63×/1.4 objective lenses. Data were acquired and processed using the PerkinElmer Volocity Verison 5.1.1 software.