Figure 5.

PBMT promotes HFSC proliferation by activating paracrine Wnt signals in SKPs

(A) Western blot detected WNT10A, WNT10B, and WNT7B protein levels in skin tissue after PBMT, n = 8 mice.

(B) Immunofluorescence detected WNT10A, WNT10B, and WNT7B expression and localization in skin tissue, n = 8 mice per group and >60 HFs per mouse.

(C) Flow cytometry was used to detect the expression of WNT10A, WNT10B, and WNT7B in the dermis and epithelium, n = 8 mice.

(D) Western blot detected the expression of WNT10A, WNT10B, and WNT7B in fibroblasts, HFSCs, and SKPs after PBMT.

(E) The expression of WNTs in SKPs was detected by immunofluorescence, n = 10 mice.

(F) qPCR analysis of the expression of WNTs genes in HFSCs after PBMT.

(G) Detection of WNT10A, WNT10B, and WNT7B in SKPs culture medium by ELISA.

(H) qPCR analysis of mRNA levels of inducibility-related genes Fgf10, Cd133, Sox2, Hey, Fgf7, Alp, and Lef1 in SKPs after PBMT.

(I) Representative spheroid image formed by SKPs with or without PBMT stimulation.

(J) Representative western blot analysis of β-CATENIN protein levels was performed in HFSCs after addition of SKPs (with or without PBMT stimulation) conditional medium.

(K) The western blot of β-CATENIN and P-GKS-3β in HFSCs that were co-cultured with SKPs as indicated: co-SKPs, co-culture with SKPs; co-P-SKPs, co-culture with PBMT-SKPs.

(L) Representative images of proliferating cell nuclear antigen (PCNA) expression in SOX9-positive HFSCs in the presence of ICG-001 (5 μM). The data represent mean ± SD, n = 3 independent replicates. ^∗p < 0.05; ^∗∗p < 0.01, ^∗∗∗p < 0.001.