Fig. 5. HDM or C48/80 upregulates the biomarkers for epithelial cell-derived type 2 cytokines and neurogenic inflammation via reactive oxygen species. (A) Normal human epidermal keratinocytes (NHEKs) were treated with HDM (10 mg/ml, left panel) or C48/80 (10 µg/ml, right panel) in the absence (−) or presence (+) of NAC, and cell extracts were harvested to perform reverse transcriptase-polymerase chain reaction (RT-PCR) experiments for epithelial cell-derived type 2 cytokines (TSLP, IL-25, IL-33) and for neurogenic inflammation (NGF, CGRP) as described in the Materials and Methods. The pictures show the typical results of three repetitive RT-PCR experiments for type 2 cytokines in HDM-treated NHEKs and C48/80-treated NHEKs. (B) Relative mRNA expression levels were semi-quantified by densitometric analysis. Left: HDM-treated NHEKs. Right: C48/80-treated NHEKs. HDM: house dust mites, NAC: N-acetyl cysteine, TSLP: thymic stromal lymphopoietin, IL: interleukin, NGF: nerve growth factor. ^**p<0.01.