Fig 2: Chromatin accessibility is significantly changed in hyperplastic KCs and reduced at the Il34 gene locus. (a, b) Venn diagrams, generated in DiffBind, show the overlap of the enriched sites identified after defining the consensus peakset (a) and after differential enrichment analysis (b). Epithelial samples from four independent experiments, each pooling cells from 6 WT or 3 K14E7 mice. (c) The association of DES with enhancers and promoters. (d, e) Gene ontology analysis of less accessible DES (d) and more accessible DES (e) that were associated with enhancer and promoter regions. (f) IGV visualisation of ATAC‐seq peaks at the Il34 gene and DES within its locus spanning 50 kb upstream and downstream. (g) IGV visualisation of Il34_DES1 (logFC = 0.53122, Adj.P = 0.00235) and Il34_DES5 (logFC = −0.69091, Adj.P = 0.00003). The red bar highlights Il34_DES5 overlapping with an upstream enhancer of Il34 highlighted by the green bar.

Fig 3: K14E7 KCs express reduced levels of Il34. (a) scRNA‐seq data 3 was used to visualise (a) sample groups (left) and Il34 expression (right) using Nebulosa, (b) cell states as t‐SNE plot, (c) Il34 expression across K14E7 and WT cell states as violin plot and (d) Csf1 expression across K14E7 and WT cell states as violin plot. (e, f) Comparison of Il34 mRNA expression (normalised by Rps6, Rpl5, eIF3a) (e) and IL34 protein expression (f) between K14E7 and WT epithelial skin cell lysates (n = 6 combined from 2 independent experiments). Unpaired t‐test with Welch's correction. ****P < 0.0001. (g) Representative of Il34‐targeted in situ hybridisation using cross‐sections of ears. The dashed line indicates the epithelium. Scale bars represent 25 μm for the top images and 5 μm for the bottom images. Il34 expression levels per cell. Unpaired t‐test with Welch's correction. ***P < 0.001. All error bars represent standard deviation (SD).