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Supplementary Fig. 5. (Fig. S5): Percentage of p63 positive nuclei, mitotic nuclei, and total cell density following PUV treatment on whole corneas.
(supplements the data in Fig. 7A–C)
Representative images of p63 positive nuclei are shown in the green fluorescence channel (Column i), mitotic nuclei (H3S10) are shown in the red fluorescence channel (Column ii), alongside corresponding Hoechst counterstain in the blue (UV) fluorescence channel (Column iii), for the time points indicated as days post treatment (dpt). (A-E) and (A′-E′) correspond to 1, 3, 5, 15, 25dpt for untreated control corneas and PUV treated corneas, as indicated. Percentage of p63 remains fairly consistent in untreated control corneas. PUV treatment reduced the percentage of p63 nuclei at 1dpt, and the nuclei appear damaged. The mitotic nuclei counts were decreased at 1dpt in PUV treated corneas, while an increase was observed in mitosis at 3 and 5dpt in the PUV treated corneas. After the initial increase, the mitotic counts declined at 15dpt to control levels, and remained lower than the mitotic counts in untreated control corneas at 25dpt. Total cell density remains fairly consistent from 1 to 15dpt in untreated control corneas, and an increase in the total cell density was observed at 25dpt. Total cell density decreased at 1dpt in PUV treated corneas, while increased total cell density was observed at 3, 5, 15, 25dpt. Scale bar in E′ column iii is 50 μm. |
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Supplementary Fig. 8. (Fig. S8): Percentage of p63 positive nuclei, mitotic nuclei, and total cell density following PUV treatment on half corneas.
(supplements the data in Fig. 7A’-7C′)
Representative images of p63 positive nuclei are shown in the green fluorescence channel (Column i), mitotic nuclei (H3S10) are shown in the red fluorescence channel (Column ii), alongside corresponding Hoechst counterstain in the blue (UV) fluorescence channel (Column iii), for the time points indicated as days post treatment (dpt). (A-E) and (A′-E′) correspond to 1, 3, 5, 15, 25dpt for untreated control half of the corneas and the PUV treated half of the corneas, as indicated. PUV treatment reduced the percentage of p63 nuclei at 1dpt in both the halves, which recovered at 15 and 25dpt. The treated half of the cornea initially showed a decrease in mitosis compared to the untreated half at 1dpt. Similar levels of mitosis were observed between the treated and untreated halves for 3, 5, 15, 25dpt, where both the halves showed an increase in mitosis at 3, 5, 25dpt. The total cell density in the treated half of the cornea, appeared similar to the untreated halves of these corneas for the respective time points, and showed an increase starting at 5dpt. Scale bar in E′ column iii is 50 μ |
