Confined migration induces heterochromatin formation and alters chromatin accessibility.

Cells that did not undergo mitosis after nuclear transit even showed a trend toward a further increase in heterochromatin levels over time ( Figures 2 D and S3 A–S3C).

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However, although statistically highly significant, the log 2 -fold changes (log2FC) of “up” peaks were typically small in magnitude, with the largest log 2 -fold changes only ≈0.4 ( Table S1 ), suggesting that any associated transcriptional changes may be relatively small and difficult to detect.

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The larger number of “down” peaks (purple) compared to “up” peaks (red) indicates an overall trend of reduction in chromatin accessibility, consistent with the global increase in H3K27me3 and H3K9me3 in the microfluidic devices and suggesting that cells migrating in high-concentration collagen matrices still exhibit local and small-scale heterochromatin formation, despite the lack of global increase.

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