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. 2025 May 13;8:745. doi: 10.1038/s42003-025-08162-0

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© The Author(s) 2025

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 licence, and indicate if you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by-nc-nd/4.0/.

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Fig. 3 — a Western blot analysis of atrial-specific CM proteins in purified versus unpurified monolayers. CM-AI + Laser purified monolayers (n = 6, Lanes 1–6) showed significant enrichment of Cx40, cTnI, and mlc2a protein expression compared to time and batch-matched non-purified monolayers. b Cx40 protein expression was significantly enriched in laser-purified hiPSC-CM monolayers compared to non-purified monolayers (0.73 ± 0.13au; n = 6 vs. 0.08 ± 0.01au; n = 3; ****P < 0.0001, unpaired t-test). c cTnI protein expression was significantly greater in laser-purified monolayers compared to non-purified hiPSC-CM atrial monolayers (0.31 ± 0.09au; n = 6 vs. 0.02 ± 0.01au; n = 3; ***P = 0.0009, unpaired t-test). d mlc2a myofilament protein expression was significantly enriched in laser-purified hiPSC-CM monolayers compared to non-purified monolayers (0.16 ± 0.08au; n = 6 vs. 0.03 ± 0.01; n = 3; *P = 0.04, unpaired t-test). e Atrial hiPSC-CM monolayer function was quantified using calcium-sensitive dye (Calbryte 520AM, 5 μM) and high-resolution optical mapping. Representative spontaneous calcium flux traces from re-plated CM-AI + laser processed monolayers (blue) and replated non-purified monolayers (black). f CM-AI + laser processed monolayers had faster spontaneous beat rate compared to non-purified monolayers (1.84 ± 0.14 Hz; n = 21 monolayers vs. 0.58 ± 0.14 Hz; n = 16 monolayers; ****P < 0.0001, unpaired t-test). g Calcium wave propagation velocity was faster in CM-AI laser processed monolayers compared to non-purified monolayers (22.0 ± 7.6 cm/s; n = 21 monolayers vs. 12.5 ± 3.5 cm/s; n = 15 monolayers; ****P < 0.0001). h Calcium transient duration 80 (CaTD80) was significantly shorter in CM-AI laser processed monolayers compared to non-purified monolayers (0.164 ± 0.02 s; n = 21 vs. 0.379 ± 0.07 s; n = 15; ****P < 0.0001, unpaired t-test).