Fig 1: GSTAs are induced in response to TF stimulation accompanied by ROS production. (A) MLO-Y4 cells were stimulated with TF for 24 h, after which total RNA was prepared and analysed for GSTA1, GSTA2 and GSTA5 expression by qPCR. Data are presented as the mean ± SD of three replicates. **P < 0.01. (B) MLO-Y4 cells were treated in the same way as in (A) and then whole-cell lysates were prepared and analysed by Western blotting using an antibody that simultaneously recognises GSTA1, GSTA2 and GSTA5. GAPDH was used as a loading control. Representative images and quantitative plots are shown. Data are presented as the mean ± SD of three replicataes. *P < 0.05. Full and non-adjusted images of immunoblots were shown in the supplementary material (Fig. S1). (C) MLO-Y4 cells were stimulated with TF for the indicated time, after which total RNA was extracted and analysed for GSTA1, GSTA2 and GSTA5 expression by qPCR. Data are presented as the mean ± SD of three replicataes. Statistical analyses were performed between control and each time point. **P < 0.01 (D) MLO-Y4 cells were stimulated with TF for 30 min in the presence of Hoechst 33258 and the ROS indicator DCFH-DA. Representative images taken under a fluorescence microscope at ×200 magnification are shown (left panel). The bars indicate 100 μm. The ratio of DCFH-DA-positive cells to all cells was determined by analysing micrographs taken at ×100 magnification (right panel). Data are presented as the mean ± SD of five replicataes. **P < 0.01.