Cells treated with vehicle (0.01% DMSO) were used as negative controls.
A shows the representative images of cells treated with BPA and
BPS (100 μM), BPAF and TBBPA (10 μM) for 24 h. The nuclei were
stained with Hoechst 33342 (blue). Double immunofluorescence in the cell lines
using the LC-3B (red) and MBD-1 (green) antibodies. Noted a clearly distinct
expression of LC3A in red vacuoles with perinuclear/nuclear localization and of
MBD-1 green that has a diffuse, throughout the cytoplasm, localization. Scale
bar = 50 μm. B-C shows the single-cell quantification of the
log-transformed total intensity of MBD1 (B) and LC3B (C) immuno-labeling 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).