Journal: Journal of agricultural and food chemistry

Article Title: Biotransformation and Epithelial Toxicity of Prenylated Phenolics from Licorice Roots ( Glycyrrhiza spp.) in 3D Apical-Out Mucus-Producing Human Enteroids.

doi: 10.1021/acs.jafc.4c03120

Figure Lengend Snippet: Figure 3. Polarity reversal in human ileal enteroids shown by microscopy imaging. Panel (A) shows brightfield microscopy (4× magnification) of enteroid polarity reversal over time from apical-in to apical-out. Scale bar = 650 μm and applies to all images. Panel (B) shows confocal microscopy (40× magnification) of apical-in (left) and apical-out (right) enteroids. Nuclei were visualized with DAPI (blue), the actin cytoskeleton in the microvilli brush border with Alexa Fluor 660 phallaoidin (white), and fucose units in the mucus layer with Ulex europaeus agglutinin I rhodamine (UAE-1) (green). ECM = extracellular protein matrix. Scale bars are 30 μm and apply to all images.

Article Snippet: Enteroids were subsequently permeabilized and blocked with 5% (w/v) BSA and 2% (v/v) Triton X-100 for 60 min. F-actin in the microvilli brush border was stained with ActinRed 555 (2 drops mL−1, ReadyProbes Reagent, Invitrogen, Waltham, MA, United States) or Alexa Fluor 660 phallaoidin (1:400, Invitrogen), fucose units in the mucus layer with Ulex europaeus agglutinin I conjugated to rhodamine (UEA-1) (1:100, Vector Laboratories, CA, United States), and nuclei with DAPI dilactate (1500 nM, Invitrogen), according to the manufacturer’s instructions.

Techniques: Microscopy, Imaging, Confocal Microscopy