Fig 1: LGR5 expression in LF/SC enteroids. (A) LGR5 RNA expression in LF/SC enteroids: cat (P11-12), dog (P27-34), cow (P18-27), horse (P29-35), pig (P48-49), sheep (P28-35), chicken (P18-28). qPCR of LGR5 and VIM expression in enteroids (Ent) and control tissue (Tis), student's t-test: *P=0.05, **P=0.005. Most LF/SC enteroids express LGR5 above the control tissue. LF/SC enteroids also have very low expression of mesenchymal marker VIM as compared to control tissue and kidney epithelial cells [cat (CRFK) and cow (MDBK)]. Enteroid values are fold change of control tissue; mean±se.m.; n.d., not detected; n=3. (B) EdU incorporation using the Click-iT EdU Alexa Fluor 647 assay in LF/SC enteroids confirms active proliferation with crypt-like regions. IFA of LF/SC enteroids probed with multi-species reactive a-EpCam (green, Alexa Fluor 488), Click-iT EdU (pink, Alexa Fluor 647), and nuclei stained with DAPI (blue). Merged EpCam and EdU images shows distinct regions of proliferation along crypt-like structures. Enteroids probed with a-EpCam (green) fail to stain pink (Alexa 647) in negative control IFAs lacking the Click-iT EdU.