Fig. 4: AREG mitigates apoptosis and enhances tube formation in endothelial cells. | Nature Communications

Fig. 4: AREG mitigates apoptosis and enhances tube formation in endothelial cells.

From: GPR174 knockdown enhances blood flow recovery in hindlimb ischemia mice model by upregulating AREG expression

Fig. 4

a Representative immunofluorescent image of CD31 (red), Ki67 (green) and DAPI (blue) and quantification of CD31+Ki67+ cells in muscle cross sections of Rag1−/− mice receiving Tregs 7 days after adoptive transplantation experiments (n = 6 for PBS → Rag1−/− mice; n = 7 for wild-type Tregs→Rag1−/− mice; n = 7 for GPR174-deficient Tregs→Rag1−/− mice). Scale bar, 50 μm. b Representative immunofluorescent images of CD31 (red), TUNEL (green), and DAPI (blue) staining and quantification of CD31+TUNEL+ cells in muscle cross sections of Rag1−/− mice receiving Tregs 7 days after adoptive transplantation experiments (n = 6 for PBS → Rag1−/− mice; n = 7 for wild-type Tregs→Rag1−/− mice; n = 7 for GPR174-deficient Tregs→Rag1−/− mice). Scale bar, 50 μm. c Representative immunofluorescent images of CD31 (red), Ki67 (green) and DAPI (blue) and quantification of CD31+Ki67+ cells in muscle cross sections of WT and Gpr174−/Y mice injection with AAV9-shAreg 14 days after HLI (n = 6 for WT mice + AAV9-shN and WT mice + AAV9-shAreg; n = 7 for Gpr174−/Y mice + AAV9-shN and Gpr174−/Y mice + AAV9-shAreg). Scale bar, 50 μm. d Representative immunofluorescent images of CD31 (red), TUNEL (green), and DAPI (blue) staining and quantification of CD31+TUNEL+ cells in muscle cross sections of WT and Gpr174−/Y mice injection with AAV9-shAreg 14 days after HLI (n = 6 for WT mice + AAV9-shN and WT mice + AAV9-shAreg; n = 7 for Gpr174−/Y mice + AAV9-shN and Gpr174−/Y mice + AAV9-shAreg). Scale bar, 50 μm. e Representative images of capillary-like structures and quantification of branch points and total tube length in mouse aortic endothelial cells stimulated with recombinant AREG for 16 h (n = 6). Scale bar, 200 μm. For all statistical plots, the data are presented as mean ± SD. One-way ANOVA with Bonferroni multiple comparisons test in (a, b). Two-way ANOVA with Bonferroni multiple comparisons test in (c, d). Two-tailed unpaired t-tests in (e). Source data are provided as a Source Data file.

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