Figure 2
PFOS (at 20 and 40 µM) perturbs the organization of actin- and microtubule (MT)-based cytoskeletons in human Sertoli cells without inducing cytotoxicity and apoptosis. (A) Human Sertoli cells at 70–80% confluency were treated with PFOS for 24 hr. Immunofluorescence analysis of human Sertoli cell F-actin (green fluorescence) and α-tubulin (green fluorescence, the building blocks of MTs) network was then performed. Cell nuclei were visualized by DAPI. Scale bar, 40 μm. PFOS induced defragmentation of actin microfilaments in which actin microfilaments no longer stretched across the Sertoli cell noted in control cells. PFOS also induced retraction of MTs from cell cytosol whereas MTs stretched across the entire cell cytosol in control cells. (B) Human Sertoli cells were exposed to PFOS at 10, 20, 40, 80 and 100 μM vs. vehicle control for 24 hr, and cell cytotoxicity was monitored by tetrazolium dye XTT-based assay. Each data point is a mean ± SD of triplicate wells from an experiment with n = 3 independent experiments and yielded similar results. **P < 0.01, 1-way ANOVA. (C) Human Sertoli cells were exposed to PFOS at 10, 20, 40, 80 and 100 μM vs. vehicle control for 24 hr, and were subjected to an apoptosis assay in which apoptotic cells displayed green fluorescence in the nuclei. Cell nuclei were visualized by DAPI. Scale bar, 120 μm.
