Figure 4. TLR3 activation alters keratinocyte differentiation and induces markers of hair follicle progenitors.

A) Cross-sectional H&E histology through healed scars treated with IL-6 (25ng) or control (PBS) at WD7. Scale bar = 100 μm. Quantification of healed epidermal thickness in healed scars after control or IL-6 addition.

B) Mean fold change in KRT1 and FLG mRNA after poly (I:C) (20μg/mL) addition to NHEK for 24 hours as determined by qRT-PCR and normalized to housekeeping gene, RPLP0.

C) Mean fold change in KRT1 mRNA with TLR3-specific or scrambled control siRNA in the presence of poly (I:C) (20μg/mL) in NHEK as determined by qRT-PCR and normalized as in 4B.

D) Mean fold change in KRT1 mRNA with TLR3-specific inhibitor or control in the presence of poly (I:C) (20μg/mL) in NHEK as determined by qRT-PCR and normalized as in 4B.

E) Mean fold change in KRT1 mRNA after IL-6 (50ng/mL) +/− cucurbitacin I in NHEK for 24 hours as determined by qRT-PCR and normalized as in 4B.

F) Mean fold change in Wnt pathway genes, Lgr5 and Lgr6, mRNA after 6 days of continuous exposure to poly (I:C) determined by qRT-PCR and normalized to housekeeping gene, RPLP0.

G) Mean fold change in KRT15 mRNA 72 hours after 24 hours of poly (I:C) (20μg/mL) treatment to NHEK as determined by qRT-PCR normalized to housekeeping gene, RPLP0.

H) Flow cytometry analysis of KRT15 protein expression of NHEKs treated as in 4G.

I) Flow cytometry analysis of CD200 protein expression of NHEKs treated as in 4G

*p < 0.05 by Student’s T-test or Single Factor ANOVA.