The JAK-STAT pathway is dynamically regulated during HF cycling

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Fig. 2 The JAK-STAT pathway is dynamically regulated during HF cycling. The JAK-STAT pathway is dynamically regulated during HF cycling. (A) Eight-week-old Rag1−/− and Tcr β/δ−/− mice were treated daily with JAK inhibitors. Mice were treated for a week, and images were taken 7 days after cessation of daily treatments. Representative pictures of three mice per genotype are shown. (B) Whole skin was harvested from mice at postnatal day 17 (D17) (catagen), day 23 (telogen), day 29 (early anagen), and day 33 (mid-anagen). Changes in gene expression were analyzed using a JAK-STAT qPCR array that includes genes involved in the JAK-STAT pathway as well as normalizing controls (Qiagen). Three mice were used per time point, each hybridized to a separate qPCR plate. Log 2 fold changes in gene expression were used to generate GEDI self-organizing maps to visualize the dynamic changes in gene expression over the hair cycle. GEDI clusters transcripts into metagenes on the basis of their similar expression pattern over time and placed them on a 5 × 6 grid. Metagenes repressed in experimental samples (D23, D29, and D33, respectively) versus control samples (D17) are in green to blue, whereas metagenes overrepresented in experimental samples are in red. The upper and lower thresholds correspond to a twofold change. Changes larger/smaller than 2× are set to maximum color. (C) The gene content of the repressed metagenes highlighted in (A) (boxed pixels) is detailed in the table. The color of boxed pixels corresponds to the color of lines in the table. (D) Skin from wild-type mice in anagen (day 30), catagen (day 42), and telogen (D50) was harvested, fixed, and stained with anti–phospho-Stat3 (anti–p-Stat3) and anti–phospho-Stat5 (anti–p-Stat5), as well as with Krt15 (a bulge marker) and P-cadherin (hair germ marker). Phospho-Stat3 is expressed in extrafollicular cells during anagen, as well as in DP cells (white arrows). In catagen, phospho-Stat3 is expressed in the DP (red arrow) and the hair germ (orange arrow). In early telogen, phospho-Stat3 is present in the hair germ cells that are closest to the DP (green arrow). Phospho-Stat5 is strongly expressed in the DP throughout the hair cycle, with expression peaking during catagen (yellow arrows). Phospho-Stat5 can also be detected in the bulge of catagen follicles (magenta arrow). Images were taken with a Zeiss confocal microscope (×40 magnification). Sivan Harel et al. Sci Adv 2015;1:e1500973 Copyright © 2015, The Authors