Figure 2 Helenalin induces reactive oxygen species and suppresses TrxR1 expression in prostate carcinoma cells. (A, B) The ROS levels were examined for 2 prostate carcinoma cell lines exposed to helenalin and/or ROS inhibitor by DCFH-DA staining. Bar=50 μM. (C, D) RT-qPCR was carried out to examine the mRNA expression of TrxR1 in DU145 or PC-3 cells exposed to helenalin and/or ROS inhibitor. (E–G) Western blot analysis was performed to determine the expression of TrxR1 protein in the 2 cells after treatment with helenalin and/or ROS inhibitor. (H–N) Western blot was utilized for detection of the expression of TXNIP, p-ERK, and p-AKT in the 2 cell lines following treatment with helenalin and/or ROS inhibitor. * P<0.05; ** P<0.01; *** P<0.001; **** P<0.0001.