Fig. 3: Scleral Wnt5a+ cluster alteration in FDM mice using RNAscope, and scleral WNT5A protein level change in FDM mice by immunofluorescence and western blot. | Nature Communications

Fig. 3: Scleral Wnt5a+ cluster alteration in FDM mice using RNAscope, and scleral WNT5A protein level change in FDM mice by immunofluorescence and western blot.

From: Decreased scleral Wnt5ahi fibroblasts exacerbate myopia progression by disrupting extracellular matrix homeostasis in mice

Fig. 3: Scleral Wnt5a+ cluster alteration in FDM mice using RNAscope, and scleral WNT5A protein level change in FDM mice by immunofluorescence and western blot.

a Locations of regionally differentiated sclera: iPPS, oPPS, and peripheral sclera. b Alteration of the number and percentage of Wnt5a+A2m+ cells and Wnt5a+ puncta in the temporal and nasal iPPS, oPPS, and peripheral sclera in normal murine eyes at age 3 weeks. n = 18 micrographs; Friedman test with Dunn’s post hoc test (two-sided) or RM-ANOVA with Bonferroni’s post hoc test (two-sided). c, d The numbers and percentages of Wnt5a+A2m+ cells and Wnt5a+ puncta in the temporal/nasal iPPS, oPPS, and peripheral sclera in NC eyes, FD-F eyes, and FD-T eyes. n = 9 micrographs for NC eyes; for FD eyes, n = 12 micrographs in FD-24h and n = 9 micrographs in FD-48h, one-way ANOVA with Bonferroni’s post hoc test or Kruskal-Wallis non-parametric test with Dunn’s post hoc test (two-sided). e The Wnt5a mRNA level in NC eyes, FD-F eyes, and FD-T eyes treated by FD-48 h or FD-2W. For NC eyes, n = 10 in FD-48 h and n = 18 in FD-2W; for FD-F eyes and FD-T eyes, n = 9; one-way ANOVA with Bonferroni’s post hoc test. WNT5A protein immunofluorescence intensity (f) and distribution (g) in the temporal/nasal iPPS, oPPS, and peripheral sclera in normal murine eyes. n = 11, Friedman tests with Dunn’s post hoc tests (two-sided). WNT5A protein immunofluorescent intensity (h) and change (i) in the temporal/nasal iPPS, oPPS, and peripheral sclera in NC eyes, FD-F eyes, and FD-T eyes. For NC eyes, n = 5 in FD-48 h and n = 6 in FD-2W; for FD-F eyes and FD-T eyes, n = 5, one-way ANOVA with Bonferroni’s post hoc test or Kruskal–Wallis non-parametric test with Dunn’s post hoc test (two-sided). Western blots (j) and densitometric quantification (k) of scleral WNT5A protein in FDM mice. n = 6, one-way ANOVA with Bonferroni’s post hoc test (two-sided). red: Wnt5a; green: A2m; blue: 4’, 6-diamidino-2-phenylindole (DAPI), iPPS the inner peripapillary sclera, oPPS the outer peripapillary sclera, peripheral the peripheral sclera. NC normal control eyes, FD-F untreated contralateral eyes of form deprivation mice, FD-T form-deprived eyes. Data are expressed as mean ± SEM.

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