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. 2016 Apr 15;7:11278. doi: 10.1038/ncomms11278

Figure 7. The level of histone H3 K4/K9/K27me3 in the skin is important for hair homeostasis.

Figure 7

(a) Experimental scheme of using demethylase inhibitors (DIs). (b,c) Western blot (b) and quantification of band intensity (c) of histone methylation from whole skin chromatin extracts of control and DI-treated mice. Band intensities were normalized to H3 signal. (d) Immunofluorescence staining of control and DI-treated (scheme 1 of (a)) hair follicle sections labelled with H3K9me3 (red) and CD34 (green). Scale bar, 20 μm. (e) Fluorescence intensity of all three marks from immunostainings were quantified for each bulge cell (CD34+). Student's t-test was used to determine statistical significance between control and DI-treated mice for each mark. (f) Immunofluorescence staining with a proliferation marker Ki67 of control and DI-treated (scheme 2 of a) skin sections. Scale bar, 20 μm. (g) The number of Ki67+ cells in f in each hair follicle were counted, and they showed reduced proliferation in DI-treated mice compared with control. The differences across the conditions were significant according to a Student's t-test. (h) Hair follicles of control and DI-treated mice were categorized in their hair cycle stages. Notice a dramatic increase in the number of telogen (Telo) hair follicles and reduction in anagen (Ana) I and II in DI-treated mice. (i) Mice were shaved, treated with DI in catagen (Cata; PD35–42; a, scheme 3), and followed long-term to monitor the hair cycle re-entry. Notice that mice in the control group show hair growth, whereas mice in DI-treated group show no growth. Avg, average.