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01). To unravel the cytoprotective mechanism of EPS, we tested whether EPS had a potential to inhibit UVB-induced ROS production in HDF. For experiments, HDF were http://www.selleck.cn/products/dabrafenib-gsk2118436.html irradiated with 50?mJ?cm?2 of UVB, and then intracellular ROS levels were checked immediately by DCF-DA-positive reactivity by confocal microscopy and cell sorting analysis. In the confocal microscopic study, DCF-DA-positive signals were not detected in nonirradiated control HDF (Fig.?3A-a). The DCF-DA-positive signals, which were strongly detected from UVB-irradiated HDF (Fig.?3A-b), were barely detected from EPS-treated, UVB-irradiated HDF (Fig.?3A-c), indicating a strong ROS-scavenging activity of EPS. To quantify the levels of intracellular ROS, FACS analysis for DCF-DA-positive http://www.selleckchem.com/products/Adriamycin.html cells was performed. When the average number of DCF-DA-positive cells of nonirradiated control HDF (Fig.?3B-a) was designated as 100%, that of UVB-irradiated HDF was increased to 2150?��?450% of control level (n?=?3, P? http://www.selleckchem.com/products/DAPT-GSI-IX.html of Nrf2 mRNA levels at 24�C72?h after transfection, compared to nontransfected control HDF (Fig.?5A). In RT-PCR and densitometric analysis, HO-1, NQO-1 and GSTpi were all downregulated in Nrf2 siRNA-transfected HDF at 24�C72?h after transfection, indicating Nrf2-dependent expression of HO-1, NQO-1 and GSTpi in HDF (Fig.?5A,B). From the result that EPS activated Nrf2 in HDF, we evaluated whether UVB irradiation could modulate Nrf2-relating enzymes. For experiments, HDF were irradiated with 50?mJ?cm?2 of UVB, and cells were harvested at 24?h of postirradiation. As for testing the effect of EPS, UVB-irradiated HDF were treated with EPS for 24?h after irradiation. In Western blot analysis, NQO-1 and GSTpi were upregulated in UVB-irradiated HDF. The UVB-induced upregulation of NQO-1 and GSTpi was further stimulated by EPS treatment (Fig.?6A). Thereafter, when UVB nonirradiated HDF were treated with EPS for 24?h, all HO-1, NQO-1 and GSTpi were upregulated in a dose-dependent manner at concentrations of 0.001�C0.1% (Fig.