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 Jun 6;9:53. doi: 10.1038/s41421-023-00543-1

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

Fig. 6 — a Schematic overview of the small molecule screening. A ~3000-compound library was screened for counteracting the differentiation tendency to presomitic mesoderm under a high CHIR dose (16 μM, 0–48 h). The differentiation efficiencies were estimated by using the CPC recognition model on day 6 bright-field images. b Preliminary small molecule screening result ordered by the predicted percentage of CPC regions on day 6. Cells of positive CTL were treated with optimal CHIR, while cells of negative CTL (DMSO) or library screening groups were treated with overdose CHIR (16 μM, 48 h). c Effect of BI-1347 (2 μM, 48 h) during preliminary screening. The heatmap and the predicted CPC regions are generated by the CPC recognition model. Scale bar, 500 μm. d Effect of BI-1347 on improving differentiation efficiency under high CHIR dose (20 μM, 48 h). Representative bright-field images and fluorescence images of cTnT on day 12 were shown. Scale bar, 1 mm. e Effect of CDK8 inhibitors (0.5 μM, 48 h) on improving the differentiation efficiency under titration of CHIR concentrations using iPS-B1 line. CHIR was added with BI-1347, AS2863619, or MSC2530818 for 0–48 h. The widths of effective CHIR concentration (percentage of cTnT⁺ cells ≥ 50%, indicates by a dashed line) were shown in the upper panel. Data are means ± SD. n = 3. Statistical significance was determined by t-test to compare the BI-1347, AS2863619, and MSC2530818 groups with the CTL group under each CHIR concentration. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; ns, not significant. f Effect of BI-1347 on improving the consistency of batch-to-batch differentiation efficiencies. Cells from different batches were treated with CHIR (6 μM, 48 h) or CHIR (12 μM, 48 h) plus BI-1347 (0.5 μM, 48 h) on the iPS-B1 line. ***P < 0.001. g Effect of BI-1347 on improving the consistency of line-to-line differentiation efficiencies. Cells from six different lines (iPS-B1, iPS-18, iPS-F, iPS-M, H9, WIBR3) were treated with CHIR (6 μM, 48 h) or CHIR (12 μM, 48 h) and BI-1347 (0.5 μM, 48 h). ****P < 0.0001. h Immunofluorescent result of CM with BI-1347 treatment (0.5 μM, 48 h) under high CHIR dose (16 μM, 48 h). Representative fluorescent images of CM with BI-1347 treatment for cTnT, MEF2C, cTNI, NKX2.5, and α-ACTININ on day 12 were shown. Scale bar, 100 μm. i Gene expression data for iPSC, iPSC-CM without BI-1347 treatment, and with BI-1347 (0.5 μM, 48 h)- treated iPSC-CM (day 12, both without metabolic purification). Heatmap shows Z-scores of log₂(FPKM + 1) normalized over multiple samples. j Action potential in iPSC-CMs with (+BI-1347, 0.5 μmol/L, 0–48 h) or without (–BI-1347) BI-1347. k I–V plots of I_Ca in iPSC-CMs with or without BI-1347. Data are expressed as means ± SEM. n = 15–16. l Fluorescent images (upper) and time courses (lower) of spontaneous calcium transients in iPSC-CMs with or without BI-1347.