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. Author manuscript; available in PMC: 2022 Dec 2.
Published in final edited form as: Nat Biomed Eng. 2021 Dec 2;5(12):1500–1516. doi: 10.1038/s41551-021-00823-9

Extended Fig. 4. Effect of stiff mechanical environments on chromatin organization in non-contractile cells in vitro.

Extended Fig. 4.

a) Embryonic cardiac cells were isolated from (E)18.5 H2b-eGFP embryo hearts and cultured on either soft (13 kPa) PDMS, stiff (140 kPa) PDMS or TCP for two or four days after which cells were stained for H3K27me3 and H3K9me3 as well as actin to distinguish non-contractile cells (NCCs) from CMs. Scales=5 µm. b) NCC nuclei were evaluated for peripheral enrichment (0=center, 1=periphery) of overall chromatin (H2b) or epigenetically marked chromatin. Gray areas indicate center and peripheral bin; SEM; n≥30 from 3 exp. c) Enrichment scores for each chromatin marker were calculated as the quotient of intensity of the peripheral bin (0.85–0.95) divided by the center bin (0.05–0.15). Substrate stiffness only minorly affected chromatin organization and enrichment of overall and H3K9me3-marked chromatin remained low while enrichment of H3K27me3-modified chromatin remained high throughout the four-day culture period. SEM; n≥30 from 3 exp.; 1W-ANOVA: * p<0.05, ** p<0.01.