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. Author manuscript; available in PMC: 2018 Nov 28.
Published in final edited form as: Cell Rep. 2018 Sep 25;24(13):3383–3392.e5. doi: 10.1016/j.celrep.2018.08.054

Figure 4. Mouse and Human Skin Exhibit Age-Dependent EZH2-Mediated SDF1 Induction.

Figure 4.

(A) Age-dependent epigenetic regulation of SDF1. Shown are H3K27me3, H3K4me3, and EZH2 chromatin immune precipitation of ear wound edge tissue at baseline and 1 week post-injury at 3 different locations on the SDF1 gene. n = 6. *p < 0.05.

(B–E) Injury-induced EZH2 increases with age. Shown are EZH2 transcript (B, n = 24), protein (C, n = 4), and immune staining (D, brown cells, black asterisks mark background staining, n = 4)(E) immunostaining quantification in ear wound edge tissue at baseline (BL) and 1 week post-injury. *p < 0.01.

(F) Pharmacologic inhibition of EZH2 (DZNep) in aged mice rescues SDF1 induction and blocks tissue regeneration. Shown are SDF1 blood serum levels and ear hole measurements. n = 3. ***p = 0.00065.

(G) Human skin exhibits age-dependent SDF1 induction. Shown are SDF1 transcript level sat baseline and 1 week post-injury in human skin of different ages. n = 5 patients < 46 years old (black bars) and n = 4 patients > 61 years old (gray bars). *p < 0.05.

(H) EZH2 knockdown rescues SDF1 induction in aged human keratinocytes. Left: western blot analysis of EZH2 and actin in WT, control (scramble) knockdown (KD), and EZH2 knockdown keratinocytes. n = 4. Right: SDF1 transcript levels at baseline (BL) and after nutrient deprivation in WT, control knockdown, and EZH2 knockdown young and aged primary human keratinocytes. n = 4. **p < 0.01.

(I) Young and aged human skin organoids exhibit age-dependent SDF1 induction. Shown are SDF1 immunostaining (brown cells) and quantification 3 days after hole punch. n = 3. *p < 0.05.

(J) EZH2 inhibition rescues SDF1 induction in aged human skin organoids. Shown are SDF1 (left) and EZH2 (center) transcript levels in young (blue bars) and aged (red bars) human skin organoids and SDF1 transcript in aged human skin organoids with and without DZNep (right). n = 6. *p < 0.02.

N, biological replicates per group. Error bars are SEM. Scale bars, 100 μM. Nuclei are counterstained blue. See also Figure S4.