Cells treated with vehicle (0.01% DMSO) were used as negative controls.
The nuclei were stained with Hoechst 33342 (blue), F-actin with Phalloidin
staining (green), and γ-H2AX with a combination of primary phosphorylated
γ–H2AX and secondary Dylight 650 conjugated antibody (red).
A shows the representative images of cells treated with BPA and
BPS (100 μM), BPAF and TBBPA (10 μM) for 24 h. Arrows indicate
dot-like structures. Scale bar = 50 μm. B-C shows the
single-cell quantification of the log-transformed total intensity of F-actin (B)
and positive γ-H2AX cells (C) of the three testicular cell types treated
with various BPA, BPS, BPAF and TBBPA concentrations for 24 and 48h. Data were
presented as mean ± SD, n = 6. Three replicates in two separate
experiments were included. Statistical analysis was conducted by 1-way ANOVA
followed by Tukey-Kramer multiple comparisons (*P < 0.05).