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. 2021 May 24;6(10):e145854. doi: 10.1172/jci.insight.145854

Figure 9. Inactivation of RAR signaling in Ret lineage at E8.5 or E10.5. (A) RarαDNLoxP/+; RETCreERT2-EYFPTandem have CreERT2 knocked into the Ret locus and ChR2-Lox-Stop-Lox-EYFP in the nearby Rosa26 locus on the same chromosome.

Figure 9

Tamoxifen activates CreERT2 to induce expression of RarαDN and EYFP. (B and C) E13.5 whole bowel from control (RarαDNLoxP/+ that lacks RETCreERT2-EYFPTandem) (B) RarαDNLoxP/+; RETCreERT2-EYFPTandem (C) after E10.5 tamoxifen treatment. ENS was visualized using TuJ1 (red) and GFP (green) antibodies. (D–I) Boxes indicate regions of esophagus (D and E), small bowel (F and G), and distal colon (H and I) that are magnified in adjacent images. Scale bars: 1000 μm (B and C) and 500 μm (D–I). (J and K) E13.5 bowel after E8.5 tamoxifen treatment. Scale bar: 1000 μm. (L and M) Regions of magnified images from RarαDNLoxP/+; RETCreERT2-EYFPTandem small bowel (L) or distal colon (M). Scale bar: 100 μm. (N and O) RET (red) and GFP (green) antibody stained E13.5 colon after E10.5 tamoxifen treatment. (P and Q) PHOX2B (green), GFP (red), and DAPI (blue) stained E13.5 colon after E10.5 tamoxifen treatment. Scale bars: 100 μm. n = 3 in each group.