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

Figure 6.

Figure 6

Immunofluorescence confocal image analysis of 3-nitrotyrosine (3-NTyr) in mitochondrial proteins in BMSCs challenged with S. epidermidis. The mitochondrial network was visualized by projections of Tom20 (red channel), a mitochondrial marker (projections of 3-NTyr appear in green). Projections of the nuclear DNA are displayed in blue (counterstaining of nuclei with Hoechst 33342). Spatial co-localization of 3-NTyr 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. (A1, B1 and C1) Projections of Tom20 and 3-NTyr in control BMSCs (A1), BMSCs challenged with S. epidermidis (B1), and BMSCs challenged with S. epidermidis in the presence of 100 μM LNIL, an inhibitor of iNOS (C1). (B4) Magnification of the selected AOI is indicated with a square in B1. (A2, B2, C2 and B5) Immunofluorescence of 3-NTyr in the images shown in A1, B1, C1 and B4 respectively. (A3, B3 and C3) Histograms representing relative intensities of immunofluorescence of 3-NTyr shown in A2, B2 and C2 respectively.