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  • Single-cell RNA sequencing identifies a migratory keratinocyte subpopulation expressing THBS1 in epidermal wound healing.

Single-cell RNA sequencing identifies a migratory keratinocyte subpopulation expressing THBS1 in epidermal wound healing.

iScience (2022-04-09)
Ratklao Siriwach, Anh Quynh Ngo, Makio Higuchi, Kentaro Arima, Satoko Sakamoto, Akira Watanabe, Shuh Narumiya, Dean Thumkeo
ABSTRACT

Keratinocyte differentiation is an intricate process that is regulated by multiple mediators. Using cultured human keratinocytes, we found that lysophosphatidic acid (LPA) induced the differentiation of a previously unsuspected keratinocyte subpopulation expressing the extracellular matrix protein, thrombospondin-1 (THBS1). This action of LPA was mediated by the RHO/ROCK-SRF signaling downstream of LPA1 and LPA5 receptors and required ERK activity. Suppression of THBS1 in vitro suggested a migratory role of THBS1+ keratinocytes. Moreover, we analyzed publicly deposited single-cell RNA sequencing dataset and identified Thbs1-expressing keratinocytes in the mouse wound skin. Immunohistochemistry analysis revealed that Thbs1+ keratinocytes were apparently differentiated from basal keratinocytes upon wounding, subsequently polarized and migrated suprabasally toward the wound front, and eventually underwent terminal differentiation in the neo-epidermis. Importantly, inhibition of Erk activity suppressed Thbs1+ keratinocyte differentiation in wound healing. Based on these findings, we suggest that THBS1+ keratinocyte is a migratory keratinocyte subpopulation that facilitates epidermal wound healing.

MATERIALS
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Brand
Product Description

Sigma-Aldrich
U0126, U0126 is a elective inhibitor of MEK-1 and MEK-2; weak inhibitor of PKC, Raf, ERK, JNK, MEKK, MKK-3, MKK-4/SEK, MKK-6, Abl, Cdk2 and Cdk4.
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Monoclonal Anti-MAP Kinase, Activated (Diphosphorylated ERK-1&2) antibody produced in mouse, clone MAPK-YT, ascites fluid
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Rho/SRF Pathway Inhibitor, CCG-1423, The Rho/SRF Pathway Inhibitor, CCG-1423, also referenced under CAS 285986-88-1, controls the biological activity of Rho/SRF. This small molecule/inhibitor is primarily used for Cell Structure applications.
Sigma-Aldrich
Anti-Ki67 Antibody, clone 1O15, ZooMAb® Rabbit Monoclonal, recombinant, expressed in HEK 293 cells