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. 2018 Jul 1;32(13-14):978–990. doi: 10.1101/gad.311910.118

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© 2018 Okano-Uchida et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 4. — Deletion of Orc1 in TGCs has no effect on endoreduplication and placental development. (A) Genotypes of the offspring from Plf-Orc1^+/f and Orc1^f/f intercrosses. (B) PCR genotyping showing Orc1 deletion in E10.5 Plf-Orc1^f/f placentae. Whole placenta sections (female genotype was Orc1^f/f) were used to extract genomic DNA. (Bottom panel) The asterisk indicates generation of a PCR product used as an internal control. (C,D) Representative images of H&E-stained (C) and Feulgen-stained (D) E10.5 control (Orc1^f/f) and Orc1-deleted (Plf-Orc1^f/f) placentae. (E) Quantification of Feulgen-stained TGCs derived from E10.5 control (Orc1^f/f and Orc1^f/Δ) and Orc1-deleted (Plf-Orc1^f/f and Plf-Orc1^f/Δ) placentae. At least 85 TGCs were counted for each placenta. (F) IF images of placentae using CRE-specific (green) and BrdU-specific (red) antibodies; slides were counterstained with Hoechst 33342 (blue). BrdU was injected 1 h prior to dissection. Arrowheads indicate TGCs stained positive for both CRE and BrdU. (G) IHC for Cyclin A2 showing comparable expression in TGCs of E10.5 control (Orc1^f/f) and Plf-Orc1^f/f placentae. Bars: C,D,F,G, 50 µm. (n) Number of mice in each group.