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. 2015 Feb 26;19(5):1133–1150. doi: 10.1111/jcmm.12518

Figure 5.

Figure 5

Immunofluorescence confocal image analysis of spatial localization of IRGM in BMSCs challenged with Staphylococcus epidermidis. (A1) Control BMSCs. Mitochondrial network was visualized by projections of Tom20 (red channel), a mitochondrial marker, projections of IRGM protein appear in green. Projections of the nuclear DNA appear in blue (counterstaining of nuclei with Hoechst 33342). Index of spatial correlation of IRGM and Tom20 was defined as R = 0.25 (see Materials and methods). (B1) Alterations in mitochondrial translocation of IRGM (green channel) observed 24 hrs after bacterial challenge. Mitochondrial network was visualized by projections of Tom20 (red channel). Index of spatial correlation of IRGM and Tom20 is defined as R = 0.36 (see Materials and methods). (A2 and B2) Relative intensities of the IRGM-immunofluorescence projections presented in the respective A1 and B1. Note: Spatial co-localization of IRGM and Tom20 appears in yellow as result of interference of green and red colours. The confocal images were taken with pinhole setup to obtain 0.3 μm Z-sections. The images shown in the figure are representative ones of analyses of three specimens obtained from three experiments.