Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2023 Apr 3;41(12):1787–1800. doi: 10.1038/s41587-023-01718-7

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2023

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/.

PMC Copyright notice

Extended Data Fig. 8 — (a) Lentiviral vector used for lineage tracing of JCF progenitors in epicardioids derived from AAVS1-CAG-FRT-flanked STOP-mKate2-HA reporter hiPSCs, consisting of the sequence encoding Tamoxifen-inducible flippase (FLP^ERT2) driven by the MAB21L2 promoter. cPPT: central polypurine tract; WPRE: woodchuck hepatitis virus posttranscriptional regulatory element. (b) (Left) Immunostaining for the HA-tag and ISL1 in epicardioids at day 9, after infection with the MAB21L2-FLP^ERT2 lentivirus at day 3 and application of hydroxtamoxifen (4-OHT) days 7-8. The arrowhead shows an exemplary HA-tag⁺ ISL1⁺ JCF cell at the outer layer. (Right) Corresponding uninfected negative control. Images are representative of 3 independent differentiations. (c,d) Experimental protocol used for lineage tracing of early MAB21L2⁺ progenitors. Epicardioids differentiated from AAVS1-FRT-flanked STOP-mKate2-HA reporter hiPSCs were transduced with a lentiviral vector encoding inducible flippase (FLP^ERT2) under the control of the MAB21L2 promoter at day 3 and treated with 4-OHT at days 4 and 5 before analysis at day 12. (c) (Left) Representative images of immunostaining for cTnT, E-cadherin (CDH1) and the HA-tag in infected organoids at day 12. Filled arrowhead: exemplary HA-tag⁺ mesothelial epicardial cell (epi), empty arrowhead: exemplary HA-tag⁺ epicardium-derived cell (EPDC), arrow: exemplary HA-tag⁺ cardiomyocyte (CM). Scale bar = 50 µm. (Right) Percentage of epi, EPDCs and CMs among HA-tag⁺ cells. Mean ± SEM; N = 12 epicardioids, 3 independent differentiations. (d) (e,f) UMAP plots showing the expression of the transcription factor (TF) GATA4 and its downstream targets in the JCF metacell cluster 14 (positively regulated: DPF3, NEBL, RP11-332H18.4; repressed: LRP2). (e) UMAP plots showing the expression of the TF TFAP2B and its downstream targets in the JCF metacell cluster 14 (positively regulated: AHNAK, CDC42EP1, CGN, ELMOD1, EPAS1, F11R, FUT8, GPC5, KCNMA1, KRT19, MXRA8, PDLIM1, PRSS23, RIMS2, SLC28A2, SPINT2, SWAP70, TGIF1, VTCN1, WNT6, WNT7B; repressed: METRN). (f).