Figure 2. Enhanced ROS decrease the expression of stemness genes in NT2 cells.

(A) Cells treated with paraquat (PQ) at the indicated concentrations were lyzed, and the protein levels of stemness markers, NANOG, OCT4 and TDGF1, were examined by Western blot. β-ACTIN was used as the loading control. Cells treated with indicated concentrations of PQ (B) or H₂O₂ (C) for two days were lyzed for total RNAs. The transcript levels of stemness genes were measured by qPCR. Bar: mean ± SD; ^*p < 0.05, ^**p < 0.01 by Student t-test. (D) Cells were treated with 50 μM paraquat or 10 nM H₂O₂ for six days and immunostained for the above stemness markers. The cells were counterstained with DAPI. Scale bar: 20 μm.