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. 2021 Nov 17;12:6645. doi: 10.1038/s41467-021-26966-6

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — a The cartoon of analysis strategy focus on DEGs in the CPM that are shared between both lineages (black). b Comparison of differentially expressed genes (DEGs) from Mesp1^Cre/+; ROSA26-GFP^f/+ (Ctrl) vs Mesp1^Cre/+; ROSA26-GFP^f/+;Tbx1^f/f (cKO) embryos at E9.5 and Tbx1^Cre/+; ROSA26-GFP^f/+ (Ctrl) vs Tbx1^Cre/f; ROSA26-GFP^f/+ (cKO) in MLPs. X axis indicates log2-fold change of Mesp1^Cre DEGs. Y axis indicates the log2-fold change of Tbx1^Cre DEGs. Each dot indicates a gene, and red indicates common DEGs with |log2-fold change | > 0.25 in both comparisons. c, d Enriched biological processes in gene ontology (GO) and related genes found in downregulated (c) or upregulated (d) genes in MLPs of Tbx1 cKO. X axis indicates adjusted P values. e Single-cell embedding graph of the CPM lineages from Mesp1^Cre Ctrl and cKO at E9.5 separated by genotype and colored by expression level (gray to blue, green, yellow, low to high). f Single-cell embedding graph of the CPM lineages from Tbx1^Cre Ctrl and cKO at E9.5 datasets separated by genotype and colored by expression level (gray to blue, green, yellow, low to high). g RNAscope in situ hybridization with EGFP (green), Aplnr (red), and Pax8 (blue) mRNA probes on the transverse section from Mesp1^Cre/+;ROSA26-GFP^f/+;Tbx1^f/+ (Ctrl) embryos (upper panel; n = 3) and Mesp1^Cre/+;ROSA26-GFP^f/+;Tbx1^f/f (cKO) embryos (lower panel; n = 3) at E9.5. Scale bar, 100 µm. The white dotted line indicates the position in higher magnification images shown in the right. Scale bar, 30 µm. h RNAscope in situ hybridization with EGFP (green), Aplnr (red), and Pax8 (blue) mRNA probes on the transverse section from Tbx1^Cre/+;ROSA26-GFP^f/+ (Ctrl) embryos (upper panel; n = 3) and Tbx1^Cre/f;ROSA26-GFP^f/+ (cKO) embryos (lower panel; n = 3) at E9.5. Scale bar, 100 µm. The white dotted line indicates the position in higher magnification images shown in the right (scale bar, 30 µm. a atrium, ao aorta, e ectoderm, nt neural tube, ph pharynx, v ventricle.