Figure 7

Sex steroids induced “non-typical” phenotypes in epidermal MCE of Xenopus embryos. A) Enhanced atp6v1a signal in MR cells treated with E2 or T. Staining: Double FISH for atp6v1a (green) and foxi1e (red), and DAPI, 2 μm stack. B) Quantification of atp6v1a enrichment based on the ratio of atp6v1a to foxi1e signal areas in single cells, d.f. = 69. C) Accumulative occurrence of “non-typical” phenotypes by condition. “Non-typical” phenotypes were not detected in controls. D) “non-typical” double itln1: tuba1a-b cell without cilia (arrow) in an EE2-treated embryo. Staining: Double FISH for itln1 (green) and tuba1a-b (red), IHC for ac-Tuba, and DAPI, 2 μm stack. E) “non-typical” double atp6v1a: tuba1a-b cell in a T-treated embryo, 2 μm stack. F,G) Orthogonal view of a normal MC cell (B) and “non-typical” double atp6v1a: tuba1a-b cell with internalized cilia in a T-treated embryo. B-D) Staining: Double FISH for atp6v1a (green) and tuba1a-b (red), IHC for cilia marker acetylated-tuba (ac-Tuba), and DAPI. H) Induction of proliferative MR and MC (dashed-line squares), and double atp6v1a: tuba1a-b cells (square). Staining: Double FISH for tuba1a-b (green) and atp6v1a (red), IHC for proliferation marker phospo-histone3 (pH3), and DAPI, 2 μm stack. A,D-H) Data from confocal microscopy imaging analysis. C-H) “non-typical” data corresponds to a screening of 64 embryos by condition. B,C) Mean ± SD, “a” = control group; statistical significance is represented in a letter code: a indicates no difference with control, all other letters indicate significant difference with the control, shared letters indicate no significant differences between the corresponding data sets. One-way ANOVA, Tukey comparison test, p < 0.05. MC = multiciliated, vMR or rMR = vesicle or ridged mitochondrion-rich, MS = mucus-secreting cells.