Skip to main content
. Author manuscript; available in PMC: 2018 Jun 8.
Published in final edited form as: Sci Immunol. 2017 Dec 8;2(18):eaao1135. doi: 10.1126/sciimmunol.aao1135

Fig. 5. Increased movement of GPR55 KO cells in association with epithelial cells.

Fig. 5

(A) Representative z-projection view of small intestinal villi of GPR55 Het (upper panel) and KO (lower panel) mice, showing CD8α-PE labeled cells (red) and cell nuclei (Hoechst, blue). Lines show tracks of labeled IELs. Images are representative from more than 6 mice in each genotype. Scale bars, 20μm. (B) Representative z-projection view of small intestinal villi of TCRδ–eGFP+ GPR55 Het (upper panel) and KO (lower panel) mice, showing eGFP (green) and cell nuclei (Hoechst, blue). Lines show tracks of GFP+ cells. Circles and ovals in A and B show regions of tracks that are localized in close association with epithelial cells. Images are representative from more than 7 mice in each genotype. Scale bars, 40μm. (C) Representative snap shots from single Z-plane movies. Left two panels from GPR55 Het and right two panels from GPR55 KO. Close association movement along epithelium observed as overlap of eGFP signal with the fluorescence signal of the epithelial cell nuclei labeled with Hoechst (circles in right upper panel). Scale bars, 10μm. (D, E) Representative multiple Z-stack-projection view (D) and single Z-plane view (E) of small intestinal villi of mixed BM chimeras of the type described in Fig. 4D. Red cells are Het (red tracks) while red and green double positive cells are KO (green tracks). Ovals in D (yellow in Het and orange in KO) show regions of tracks that are localized in close association with epithelial cells. Dashed white lines in E indicate the boundaries of the Hoechst labeled epithelial cell nuclei (ECN) and the peri-basement membrane space (PMS) used for the quantitation. Images are representative from 6 mice. Scale bars, 20μm. (F) Speed of GPR55 Het and KO γδT IELs in mixed BM chimeras. (G, H) Percentage of time in close association with epithelial cells and time of each association (G), and frequency of lateral intercellular space (LIS)-probing, and time of each probing (H) of GPR55 Het and KO cells in mixed BM chimeras. Combined data from 6 movies from 6 different chimeric mice. (I) Induction of Pkr and Usp18 mRNA in the small intestine of GPR55 Het and KO mice treated with anti-CD3 Ab for 3 hours, plotted relative to Hprt. Combined data from two independent experiments (n=9 in each group).***p<0.001, **p<0.01, * p<0.05, ‘n.s.’ p>0.05 by Student’s t-test.