Figure 2.

Fluorescence analysis of brush border localization in 3D spheroids/organoids from human, mouse, pig and chicken origin. A new spheroid passage was cultured for 3 days (5 days for human) in WERN medium and then spheroids were either maintained in WERN medium or differentiated in ERN medium for further 3 days (human for 5 days). As a control, 3/5 day spheroids in WERN were also assessed in parallel. Subsequently spheroids/organoids were fixed and F-actin in the microvilli of the luminal brush border was visualized by staining with Phalloidin iFluor-488 (green). Cell nuclei were stained with DAPI (blue). Note, video composition of z-stacks are also provided as Supplementary Material. Scale bars represent 50 µm. All experiments were performed twice with similar results.