Fig. 5
Quantification of immune cells in cryosections of damaged retina. A Quantification of L-plastin+ immune cells per 1000 μm² of inner retina (5-μm-thick sections) from saline or ouabain injected eyes at 24, 48, and 72 hpi, representative images in Fig. 3G–L). For details, see the “Quantification of Immune Cells in Retinal Cryosections (24–72 h post-injection)” section. Regions depicted by dotted white lines in Fig. 3G–L are representative regions used for quantification. DAPI+ nuclei surrounded by L-plastin+ signal were counted within the region of interest and normalized per 1000 μm². p values comparing densities between 24 and 48 hpi and 48–72 hpi ouabain injection are shown on the graph; asterisks indicate the following p values comparing ouabain to saline injected at indicated time points: *p = 0.012, **p = 7.84 × 10− 12, ***p = 1.21 × 10− 12. B Since thickness of the INL changes due to ouabain-induced lesion, the number of L-plastin+ cells is also reported per 100 μm of curvi-linear distance of retina using a line that follows parallel to the basal border of the ONL. p values comparing densities between 24 and 48 hpi and 48–72 hpi ouabain injection are shown on the graph; asterisks indicate the following p values comparing ouabain injected to saline injected at indicated time points: *p = 3.84 × 10− 6, **p = 8.48 × 10− 9, ***p = 5.5 × 10− 10. C PCNA+, DAPI+ nuclei surrounded by L-plastin+ signal quantified in retinal cryosections from saline or ouabain injected eyes at 24, 48, and 72 h hpi. Graph shows the fraction of L-plastin+ cell bodies with nuclear co-label of DAPI and PCNA. Only images from central and central-peripheral retina were quantified, and only L-plastin+ cell bodies within retinal tissue boundaries were counted. Error bars indicate standard deviation. *p = 0.0005, **p = 1.01 × 10− 12, comparing ouabain injected to saline injected at indicated time points