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. 2016 Jun 23;2(5):648–662.e8. doi: 10.1016/j.jcmgh.2016.06.002

Figure 4.

Figure 4

LGR4 and LGR5 are required for endoderm differentiation. (A) qRT-PCR showing mRNA abundance in all samples for LGR4, LGR5, FOXA2, and SOX17. h9–shRNA–scrambled, h9–shRNA–LGR4 knockdown, h9–shRNA–LGR5 knockdown, and h9–shRNA–LGR4/5 knockdown lines all were treated for 3 days with ACTA to induce endoderm formation. One-way analysis of variance was used for statistical analysis. (B) h9–shRNA–scrambled, h9–shRNA–LGR4 knockdown, h9–shRNA–LGR5 knockdown, and h9–shRNA–LGR4/5 knockdown lines all were treated for 3 days with ACTA to induce endoderm formation, and immunostained for FOXA2 (red) and SOX17 (green). Nuclei were stained with DAPI (blue). Scale bars represent 200 μm. (C) Cell staining shown in panel B was quantitated. Each color group, corresponding to the percentage of FOXA2+ cells (red), the percentage of SOX17+ cells (green), and the percentage of double-positive cells (blue) was compared using 1-way analysis of variance. Dissimilar letters in a specific color group indicate significantly different values (P ≤ .05), whereas similar letters in a color group indicate no difference. DAPI, 4',6-diamidino-2-phenylindole. *P ≤ .05, **P ≤ .01, ***P ≤ .001, and ****P ≤ .0001.