Hair follicle stem cells regulate retinoid metabolism to maintain the self-renewal niche for melanocyte stem cells
- Peking Union Medical College, China
- National Institute of Biological Sciences, China
- Shanghai Institutes for Biological Sciences, University of Chinese Academy of Sciences, China
- Tsinghua University, China
Figures
Figure 1 with 1 supplement
Ablation of Notch pathway effector RBP-J in bulge epithelial cells causes McSCs ectopic differentiation in the niche.
(A) Representative immunofluorescence images and quantification of Tamoxifen induced Sox9-CreER::Ai14 mice indicating efficient labeling of bulge epithelial cells but not McSCs. DCT is a melanocyte …
Figure 1—figure supplement 1
HF phenotype in Sox9-CreER::RBP-J cKO mice.
(A) Representative tail skin immunofluorescence images of Sox9 in WT and Sox9-CreER::RBP-J cKO HFs at P18 anagen. Note the expression pattern of Sox9 is similar in cKO compared to WT bulge. (B) …
Figure 2 with 1 supplement
Loss of RBP-J specifically in HFSC causes hair cycle dependent McSCs ectopic differentiation in the shared niche.
(A) Schematic diagram of CRISPR/Cas9-mediated knockin of CreER in the 3’ UTR of Krt6a locus. sgRNA targets downstream of Krt6a stop codon. Two pair of indicated primers spanning the junctions of …
Figure 2—figure supplement 1
HF phenotype in Krt6-CreER::RBP-J cKO mice.
(A) Representative fluorescent image and quantification of Krt6-CreER labeling pattern in telogen HFs. (B) FACS profile and quantification of HFSC labeling efficiency using Krt6-CreER::Ai6 mice. …
Figure 3 with 2 supplements
RBP-J functions as a repressor in HFSC and downregulates retinoid metabolic process genes.
(A) Representative immunofluorescence images of RBP-J demonstrate broad expression pattern in HF epithelial cells at all stages of hair cycle from morphogenesis to adult. Krt14 is a basal epithelial …
Figure 3—figure supplement 1
FACS sorting strategy for HFSCs, CPL cells and matrix cells.
(A) Schematic diagram of P13 anagen HF used for FACS analysis. HFSCs marked in red reside in upper HF just beneath the sebaceous gland. Matrix cells containing terminally differentiated cells marked …
Figure 3—figure supplement 2
Signal pathways that do not affect McSCs ectopic differentiation triggered by RBP-J loss in HFSCs.
(A) Representative non-specific lipid marker Oil Red O staining in the WT and Krt6-CreER::RBP-J cKO HFs at telogen. Follicles are counter-stained by hematoxylin. Note the normal and restricted red …
Figure 4
Chromatin accessibility change after loss of RBP-J in HFSCs.
(A) ATAC-seq signal comparison between FACS isolated HFSCs from WT and Krt6-CreER::RBP-J cKO HFs at D2 after wax. Each dot represents an ATAC-seq peak. Regions with more accessibility in cKO than in …
Figure 5 with 1 supplement
Increased RA from RBP-J null HFSCs causes McSCs to differentiate abnormally in the niche.
(A) Schematic diagram of retinoic acid synthesis inhibitor treatment experiment. Tamoxifen was injected on P50-52 at telogen to induce ablation of RBP-J in Krt6-CreER::RBP-J mice, then CPL cells …
Figure 5—figure supplement 1
FACS sorting strategy, validation and quantification of McSCs and other cell types.
(A) FACS profile of HG cells and qPCR validation of marker genes. Sca1 was used to exclude epidermis. CD34 was used to exclude HFSC and HG cells were gated as Sca1-, CD34-, Pcadhigh. (B) FACS …
Figure 6 with 1 supplement
RA from cKO HFSCs enhances c-Kit protein level on McSCs that leads to abnormal differentiation.
(A) Schematic diagram, FACS profiles and quantification of c-Kit+ McSCs in telogen HFs from WT and Krt6-CreER::RBP-J cKO mice after indicated treatments. Tamoxifen was injected on P50-52 at telogen …
Figure 6—figure supplement 1
Quantification of c-Kit level on McSCs and numbers of melanocytes.
(A) Percentage of c-Kit+ cells quantified by FACS under indicated treatments in WT and Sox9-CreER::RBP-J skin. (B) Schematic diagram, FACS profiles and quantification of c-Kithigh/c-Kit+ McSCs in …
Figure 7 with 1 supplement
Hair cycle dependent increase of HFSC secreted SCF induces aberrant differentiation of RA sensitized McSC in the shared niche.
(A) Representative immunofluorescence images and quantification of pERK+ McSCs in WT and Krt6-CreER::RBP-J cKO HFs. (B) Representative dorsal skin Masson-Fontana staining images and quantification …
Figure 7—figure supplement 1
Effect of Rapamycin or RA treatments on McSCs.
(A) Schematic diagram, representative dorsal skin Masson-Fontana staining images and quantification of ectopic pigmentation in the bulge of WT and Krt6-CreER::RBP-J HFs after treatment with vehicle …
Additional files
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Supplementary file 1
Genes Up-regulated in RBP-J cKO versus WT dorsal HFSCs.
RNA-seq analysis of WT and cKO HFSCs isolated from Krt6-CreER::RBP-J mice at D2 after wax to profile their transcriptome difference. Since RBP-J serves as a repressor in HFSCs, genes regulated >1.5 fold (p<0.05) in cKO compared to WT HFSCs were mainly focused.
- https://cdn.elifesciences.org/articles/52712/elife-52712-supp1-v2.xlsx
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Supplementary file 2
Primer sequences used.
- https://cdn.elifesciences.org/articles/52712/elife-52712-supp2-v2.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/52712/elife-52712-transrepform-v2.pdf
