Figure 2 of Li, Mol Vis 2020; 26:603-612.

Figure 2. Confocal fluorescent images show CP49 protein distribution in lens fibers at zone I, zone II, and zone III from the 37 °C, 25 °C, and 4 °C treated B6WT lenses. The lens cross-sectional samples were immunolabeled with anti-CP49 antibody (red), fluorescein (FITC)-phalloidin (green), and 4’,6-diamidino-2-phenylindole (DAPI; blue). The left panels are CP49 alone, and the right panels are merged with F-actin and nuclei. A, B, and C represent an area (50 μm × 50 μm) of fibers located in 0–50, 325–375, and 425–475 μm from the lens equatorial surface, respectively (see the locations of zones I, II, and III in A, B, and C, respectively, illustrated in Figure 5). D: Enlarged images of the white-box regions in zone III (C); white arrows indicate CP49 accumulation. E, F, and G: Bar graphs show CP49 staining intensity ratios between fiber cell boundaries versus the cytosol in zones I, II, and III that were quantitatively measured and compared as mean ± standard deviation (SD; n = 3, per sample group). E: Among three different temperatures, there was no statistically significant difference in the CP49 distribution in zone I. F: However, the CP49 boundary/cytosolic intensity ratio in the 4 °C treated fibers was statistically significantly higher than those in the 37 °C and 25 °C treated fibers in zone II (***p≤0.001), and the 4 °C and 25 °C treated fibers showed statistically significantly higher CP49 ratios than the 37 °C treated fibers in zone III (G, **p≤0.01 or ***p≤0.001). Scale bar: AC, 10 μm; D, 4 μm.