Figure 2
Characterisation of Lrig1-expressing cells in gastric epithelium. (A) Overview of the dissection strategy used to isolate corpus and pylorus from mouse stomach. The indicated margin between the two regions was discarded in order to avoid cross-contamination between compartments. (B) Flow cytometry analysis of mouse stomach epithelium showing differences in expression of Lrig1-eGFP and the separation strategy for FACS isolation of eGFPlow and eGFPhigh cells. (C,D) qPCR gene expression analysis epithelial cells isolated by FACS from corpus and pylorus showing enrichment of different lineage markers in analysed cell populations. Expression levels are relative to GAPDH. (E) Expression of Lrig1 in the basal layer of mouse forestomach epithelium, showing overlapping pattern with keratin 14 (K14) and proliferating cells marked by EdU, but not keratin 10 (K10). (F) Co-localisation of Lrig1 and Lgr5-eGFP in the glandular epithelium of the pylorus. (G) Co-expression of stomach cell lineage markers, H+/K+ ATPase (Atp4a) and chromogranin A (ChgA) with Lrig1. (H) Co-localisation of Lrig1 expression with actively replicating visualised by staining of incorporated EdU. White arrowheads in (G,H) indicate cells co-localisation of markers with Lrig1. Empty arrowheads point to cells showing expression of a marker without simultaneous co-expression of Lrig1. Scale bars: E −25 μm, (F–H) −50 μm. *P ≤ 0.05.
