Fig. 4

Fig. 4 Confinement-mediated stabilization of HMGB2 increases chromatin accessibility at neuronal loci.

a, Representative stills from time-lapse imaging of A375 cells expressing HMGB2–GFP and subjected to FRAP. The yellow dashed region indicates the photobleached area. Time is relative to photobleaching. Scale bars, 10 µm. b,c, FRAP recovery curves for HMGB2–GFP in unconfined (b) and confined (c) cells. Each curve represents fluorescence recovery within the area photobleached on a single cell. d,e, Representative plots showing a two-component exponential equation fit to HMGB2–GFP fluorescence recovery curves in unconfined (d) and confined (e) cells. f, Relative proportion of slow-diffusing HMGB2–GFP. Horizontal lines, mean; box, s.e.m.; vertical lines, s.d. P value is indicated (two-sample t-test; two-sided). Unconfined, n = 45 cells; confined, n = 54 cells (b–f). g, Volcano plot of differentially expressed peaks upon HMGB2^OE. P-value cut-off, 0.05; fold change cut-off, log₂(1.25) and log₂(−1.25). h, Top 10 enriched GO: Biological Process (BP) pathways from genes mapped to open chromatin loci upon HMGB2^OE. P values are indicated (two-sided hypergeometric test with Benjamini–Hochberg correction) (g,h). i,j, Tornado plots showing chromatin accessibility at loci linked to genes from the ‘Lee neural crest stem cell up’ (i) and ‘regulation of nervous system development’ (j) pathways. k, Homer de novo motif analysis of transcription factor motifs in open chromatin regions in promoter regions of genes upon HMGB2^OE. P values are indicated (binomial test).

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