FIG 5.

Transcriptional gene expression and immune localization of cryptdins in the ileum of Muc2^+/+ and Muc2^−/− littermates inoculated with E. histolytica. (A) mRNA expression levels of the cryptdin 5, cryptdin 22, and cryptdin-related sequence 1 genes in the ileum of mice inoculated with E. histolytica or PBS, as described in the legend of Fig. 1, were quantified by qRT-PCR. Data are expressed as fold changes relative to the value for the housekeeping gene Gapdh. Means ± SE are shown (n = 3 independent experiments run in triplicate). (B) Ileal sections were immunoblotted with anti-Crp1 (red) antibody and quantified by immunofluorescence microscopy. Nuclei were stained with DAPI (blue). IgG was used as an antibody control. The fluorescence intensity was quantified by using ImageJ software and averaged over 10 random fields of view in two to three independent slides per animal and is represented as MFI (histogram) normalized to the area of the field of view. Means ± SE are shown (n = 2 independent experiments run in triplicate). P values for all significant comparisons (*, P < 0.05; **, P < 0.01) are represented (one-way ANOVA followed by Bonferroni posttests). Images are from 1 of 4 independent experiments.