Figure 5. The canonical Glp2r is expressed in hepatic stellate cells.

(A) The expression of human GLP2R and GAPDH transcripts was assessed by RT-PCR in total RNA in human livers obtained from 3 indicated commercial sources and in RNA from a human GLP2R expressing human colon cancer epithelial cell line (DLD-1) as a positive control. For reverse transcription PCR of GLP2R we used primers that amplify a 1.687-kb product encompassing the entire GLP2R open reading frame. PCR products were analyzed by agarose gel electrophoresis followed by SYBR green staining (top and bottom) or by Southern blot with an internal GLP2R ³²-P–labeled oligonucleotide (middle). RT, reverse transcriptase. (B) Levels of Glp2r mRNA, relative to Ppia, in whole liver (Liv, n = 6), isolated hepatocytes (Hep, n = 10), and nonparenchymal liver cells (NPC, n = 4). (C) Hepatic stellate cells (HSC; n = 3) purified by centrifugation and flow cytometry. (D) mRNA abundance, relative to Ppia, of HSC (Col1a1, Pdgfrb) and nonstellate cell (Spic, Cd11b) markers in purified HSCs and whole liver. (E) Microscopy image demonstrating lipid vacuole autofluorescence in purified HSC. Scale bar: 20 μm. (F) Glp2r mRNA abundance, relative to Ppia, in jejunum (Jej), liver (Liv), pellet collected after Nycodenz gradient isolation of HSC (Pel), purified HSC fraction gated by size and positive violet autofluorescence (HSC), cells positive for violet and FITC autofluorescence that correspond with HSCs mixed with contaminant cells (FITC), cells gated negative to violet autofluorescence (Vio-), and cells outside size gate (Size-). (G) Full-length mouse Glp2r expression in jejunum, whole liver, hepatocytes, and isolated HSCs. Glp2r mRNA transcripts were amplified by reverse transcription PCR with primers that amplify a 1.65-kb product encompassing the entire Glp2r open reading frame. PCR products were analyzed by agarose gel electrophoresis followed by SYBR green staining (top and bottom). Southern blotting with an internal Glp2r ³²-P–labeled oligonucleotide (middle) was used to detect the PCR product. (H) mRNA abundance, relative to Ppia, of immediate-early genes used as markers of GLP-2 action in isolated HSCs. Hepatic stellate cells were purified from 18 C57BL/6 WT mice at 4–6 months old. Cells, combined from 1–4 mice per data point, were incubated for 20 minutes with either 10% FBS, GLP-2 (50 nM), or saline (Veh). Data are presented as the mean ± SEM. *P < 0.05, **P < 0.01, using ratio-paired t test.