Fig. 4

CD68 pixel clusters move away from the cell membrane from 48 h to 7 days after injury. GLCM data are consistent with increased lysosome clustering at 48 h than 7 days, as detailed by five-color-coded ramp images (a) showing spreading of low GL values (blue-green) at 7 days (white outline representing CD11b inner boundary, asterisks indicate nuclei, scale bar = 10 μm). After manual tracing of the inner CD11b boundary (white outline, b, scale bar = 1000 nm), CD68 signal intensity was analyzed at different distances (100, 300, 500, 700, and > 700 nm from the traced boundary, b′). CD68 integrated density (ID) fraction was higher within 700 nm and conversely lower at > 700 nm from the membrane inner interface at 48 h than at 7 days (c). Individual cell values with bars at mean, n = 20 cells, normal distribution by Kolmorgov-Smirnov analysis, two-way ANOVA F(4, 152) = 5.826, followed by Sidak’s post-hoc test,***p < 0.01