Different intracellular localization of CXCL4 and CXCL4L1 in HEK transfectants. (A) Strong expression of CXCL4L1 (green) in the Golgi of CXCL4L1-HEK transfectants, as shown by costaining with the Golgi marker Bodipy (red). Colocalization is shown in yellow (bar represents 20 μm). (B) Virtually absent colocalization between the DCG marker VAMP-1/2/3 (red) and CXCL4L1 (green) in CXCL4L1-HEK transfectants. Colocalization is shown in yellow (bar represents 20 μm). (C) Absence of any effect of PMA treatment in CXCL4L1-HEK transfectants on CXCL4L1 (green) distribution and localization (bar represents 20 μm). (D) Marginal colocalization of CXCL4 (green) with the Golgi marker Bodipy (red) in CXCL4-HEK transfectants. Colocalization is shown in yellow (bar represents 20 μm). (E) Localization of CXCL4 (green) within DCGs in CXCL4-HEK transfectants, as demonstrated by strong costaining with the DCG marker VAMP-1/2/3. Colocalization is shown in yellow (bar represents 20 μm). (F) Treatment of CXCL4-HEK transfectants with PMA for 6 hours induces degranulation of CXCL4-containing DCGs. Colocalization of CXCL4 (green) within DCGs of CXCL4-HEK transfectants, as stained with VAMP-1/2/3 is shown in yellow (bar represents 20 μm). Images were acquired using an LSM 510 Meta confocal microscope (Zeiss) equipped with a 20×/0.50 Plan-Neofluor objective lens (Zeiss). LSM 510 Meta confocal microscope software version 3.0 (Zeiss) was used to capture all the images.