Fig. 3

Other GKS proteins co-localize with lysosomes in Irgm1 ^−/− cells. a Wild type (WT), Irgm1 ^−/−, Irgm3 ^−/−, and Irgm1/Irgm3 ^−/− mouse embryonic fibroblasts (MEFs) were induced with 200 U/mL IFN-γ for 24 hours. Cells were fixed and stained with anti-Irgb6 antiserum (141/3) and anti-LAMP1 antibody. Representative microscopic images of Irgb6 and lysosome co-localization are shown. Arrows point at the Irgb6 structures magnified at the end of each panel in the following array: upper left: Irgb6, upper right: LAMP1, lower left: merge, lower right: phase contrast. Scale bars represent 10 μM. b Quantification of 3a, S3A and S3B, showing percent of Irgb6, Irgb10, and Irgd structures co-localizing with LAMP1; 50 cells per sample were quantified and results of two independent experiments are shown. c Irgm1 ^−/− MEFs were induced with 200 U/mL IFN-γ for 24 hours, fixed and stained for Irga6 (10D7), Irgb6 (141/3) and LAMP1; Irgd, Irga6 (10D7) and LAMP1; Irga6 (10D7), Irgb10 and LAMP1; or Irgb6 (A20), Irgb10 and LAMP1. Representative microscopic images of GKS structures and lysosome co-localization are shown. Arrows point at the GKS structures which are magnified at the end of each panel in the following array: upper left: first GKS protein, upper right: second GKS protein, lower left: LAMP1, lower right: merge. Scale bar represents 10 μM