Journal: bioRxiv

Article Title: Lgr5+ telocytes are a signaling hub at the intestinal villus tip

doi: 10.1101/850909

Figure Lengend Snippet: a) Grem1 mRNAs (green dots, green arrows mark positive cells) are expressed in crypt telocytes and not in VTTs. Scale bar – 5 µm. b) Angptl4 (green dots) are expressed more highly in VTTs compared to crypt telocytes. Scale bar – 5 µm. Red dots are Lgr5 mRNAs (red arrows mark positive cells), blue is PDGFRa antibody staining. c) Quantification of smFISH validations for crypt telocytes and VTT markers. ** denote pvalue<10 −4 from two sided Wilcoxon ranksum test, c – crypt, vt – villus tip. Boxes show 25-75 percentiles of the smFISH expression, vertical lines are medians. d) Enteric mesothelial fibroblasts are scattered throughout the crypt-villus axis. Grey dots are mRNAs of Rgs5. blue is PDGFRa antibody staining. Scale bars – 20 µm, insets scale bar – 10 µm e) Myofibroblasts are scattered throughout the crypt-villus axis and localized away from the epithelial layer. Grey dots are mRNAs of Acta2. Blue is PDGFRa antibody staining. Scale bars – 20 µm, insets scale bar – 10 µm. White arrows mark positive cells in d) and e). White dashed line mark crypt borders.

Article Snippet: For PDGFRa immunofluorescence staining, goat anti mouse PDGFRa was used as first antibody (AF1062 R&D systems) at a concentration of 8 μg/μl in the smFISH hybridization buffer, Alexa fluor 488 conjugated Donkey anti goat (705-545-147 Jackson laboratories, 1:400) was used as secondary antibody.

Techniques: Staining, Expressing