Fig. 4. Effect of SP in production of inflammatory cytokines and hair growth-related factors in outer root sheath cells. (A) Detection of neurokinin-1 receptor (NK-1R) in epidermal keratinocytes (KC) and outer root sheath (ORS) cells, using western blot analysis. (B) Treatment of ORS cells with recombinant SP (1 µM) for 2 hours. Assessment of the mRNA expression of inflammatory cytokines by quantitative PCR (q-PCR) (n=6). (C) Treatment of ORS cells with recombinant SP (1 µM) for 24 hours. Assessment of the secretion of inflammatory cytokines by ELISA (n=3). (D) Treatment of ORS cells with recombinant SP (1 µM) for 30 minutes. Determination of protein level of phospho-p65 by western blot analysis. (E) Treatment of ORS cells with recombinant SP (1 µM) for 24 hours. Assessment of the mRNA expression of vascular endothelial growth factor (VEGF) and keratinocyte growth factor (KGF) by q-PCR (n=6). IL: interleukin, CTL: control, SP: substance P. ^*Statistically significant (p<0.05).