Figure 5.

Immunofluorescence analysis of ODMs from human, mouse, pig and chicken origin. (A) Orthogonal stacks for brush border protein EZR, enterocyte marker NHE3, and SOX9. ODMs were cultured as described and fixed after 10 (human), 12 (mouse), 6 (pig) and 15 (chicken) days. Note, NHE3 analysis for pig was excluded as suitability of the antibody for IFA staining in this species is questionable (see Supplementary Figure 6). Suitability of the SOX9 antibody for IFA staining for pig is also questionable, however, the analysis was included as the antibody target sequences are highly similar between species (see Supplementary Figure 6). Representative fluorescent orthogonal stacks of ODMs are shown. Scale bars represent 20 µm. (B) Representative confocal image projections of ODMs infected for 48 h with T. gondii (distinguished by its GFP-tagged green tubular mitochondria) in TYI-S-33 medium. Scale bar 10µm. All experiments were performed at least twice with similar results.