Cell proliferation and goblet cell number in the crypts of the intestine. (A) Immunohistochemical staining against PCNA and nuclear counterstain with DAPI. From left to right: fish underwent 21 days of DMSO, 21 days of prednisolone, 17 days of DMSO followed by 4 days of prednisolone and 17 days of prednisolone followed by 4 days of DMSO treatment. White asterisks indicate goblet cells and white arrows indicate PCNA+ cells in the crypts. Scalebar 50 µm. Sample “Pred 17d DMSO 4d” showed increased autofluorescence. (B) Quantification of PCNA+ cells per crypt in the intestine. Data are mean ± SD of PCNA+ cells per crypt for the treatment conditions 21 days of DMSO, 21 days of prednisolone, 17 days of DMSO followed by 4 days of prednisolone and 17 days of prednisolone followed by 4 days of DMSO. Each dot represents one biological replicate. Non-parametric testing because of non-normal distribution of the data. Statistical significance was tested by post-hoc Dunn’s multiple comparison after Kruskal-Wallis test. n=5 (4 females, 1 male in DMSO 21d, DMSO 17d Pred 4d, Pred 17 d DMSO 4d; 5 females in Pred 21d) in all groups with 3 sections per individual. (C) Quantification of goblet cells per intestinal crypt. Data are mean ± SD of goblet cells per crypt for the treatment conditions 21 days of DMSO, 21 days of prednisolone, 17 days of DMSO followed by 4 days of prednisolone and 17 days of prednisolone followed by 4 days of DMSO. Each dot represents one biological replicate. Parametric testing because of normal distribution of the data. Statistical significance was tested by post-hoc Dunnett’s multiple comparison after one-way ANOVA. n=5 (4 females, 1 male in DMSO 21d, DMSO 17d Pred 4d, Pred 17 d DMSO 4d; 5 females in Pred 21d) in all groups with 8 sections per individual.
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