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Fig. 7 | BMC Biology

Fig. 7

From: Loss of the interferon-γ-inducible regulatory immunity-related GTPase (IRG), Irgm1, causes activation of effector IRG proteins on lysosomes, damaging lysosomal function and predicting the dramatic susceptibility of Irgm1-deficient mice to infection

Fig. 7

Lysosomal processing is impaired in IFN-γ-induced Irgm1 −/− mouse embryonic fibroblasts (MEFs). a Irgm1 −/− MEFs were transfected with EGFP-Irga6 and simultaneously induced with 200 U/mL IFN-γ. After 24 hours, non-fixed cells were incubated with 50 nM Lysotracker for 15 minutes and pictures of live cells were taken. Representative microscopic images of Irga6 and Lysotracker co-localization are shown. Arrows point at the Irga6 structures magnified at the end of each panel in the following array: upper left: Irga6, upper right: Lysotracker, lower left: merge, lower right: phase contrast. Scale bars represent 10 μM. b Quantification of 7a showing the number of Irga6 structures co-localizing with Lysotracker. 25 cells per sample were quantified and the means of three independent experiments ± standard deviation are shown. c Wild type (WT), Irgm1 −/−, Irgm3 −/−, and Irgm1/Irgm3 −/− MEFs were induced with 200 U/mL IFN-γ for 27 hours, 200 nM μg/mL Bafilomycin A1 for 4 hours, or left untreated. After 24 hours, non-fixed cells were incubated with 10 μg/mL DQ-BSA, incubated for 3 hours, and images of live cells were taken. Representative microscopic images of DQ-BSA staining are shown. Scale bars represent 10 μM. d Quantification of Fig. 7c showing the average numbers of DQ-positive structures per cells; 20 cells per sample were evaluated and results of three independent experiments ± standard deviation are shown. Differences were tested for statistical significance using the Mann-Whitney test (NS P > 0.03)

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