Extended Data Fig. 7: miR-31 promotes HFSCs transepidermal differentiation and aging by activating MAPK/ERK.
From: A stress-induced miR-31–CLOCK–ERK pathway is a key driver and therapeutic target for skin aging
a, Representative FACS sorting profiles of CD49fhiCD34 + HFSCs (box) from P52 WT/DTG littermates (+Dox since P48 and waxed at P50 as indicated in the top diagram). % numbers indicate percentage of the HFSCs among the CD49f + epithelial cells. b, Volcano plot of gene expression changes between backskin HFSCs from WT/DTG littermates based on RNA-seq analysis. Each data point represents a gene. Red points indicate significantly changed genes (Padj < 0.01, Padj values were two-side and multiple comparison adjusted P values calculated by Deseq2). c, Western blots of cultured MK or NHEK cells stably expressing miR-31 or scrambled control miRNA (Scr) as indicated in Fig. 5b. Left labels: primary antibody used. Numbers indicate relative quantification (vs MK Scr for left, vs NHEK Scr for right). Green signals were first normalized to reference protein β-actin (ACT). d, Left: Western-blots of EDTA isolated backskin epithelium from young (8 weeks)/old (22 months) mice. Numbers indicate relative quantification of their above green bands. Right: Quantification of the left pERK bands. n = 3 replicated wells. e, Western blots of ERK/pERK in backskin epithelium of WT/DTG littermates treated with Trametinib (+T) or solvent control (+S). Numbers indicate relative quantification (vs WT + S). f, Western blots of ERK/pERK in Scr or miR-31 overexpressing MK cells treated with Trametinib (+T) or solvent control (+S). Numbers indicate relative quantification (vs Scr+S). g, Representative FACS profiles for backskin epithelium HFSC abundance quantification in Fig. 5g. h, Representative backskin photographs of Dox induced, waxed, and sch772984 (+sch) or solvent control (+S) treated WT/DTG littermates at d15 post waxing. Top diagram: treatment procedures. Scale bars=1 cm. For all relevant figures, data are represented as mean ± SEM. P values were calculated with two tail t-test.
