Fig. 10

Cadmium-induced Sertoli cell injury that affects the cytoskeletal organization of microtubules is mediated through p38 MAPK activation. A A study by immunofluorescence microscopy to assess the organization of microtubule cytoskeleton network across the Sertoli cell cytoplasm. Pre-treatment of Sertoli cell epithelium on day 3 with doramapimod (10 µM), a p38 MAPK inhibitor, was able to block the cadmium-induced Sertoli cell injury by causing dis-organization of the MT network across the cell cytoplasm. CdCl₂ dissolved in 0.9% saline (v/v, in Milli Q water) was used at 1 µM. Results shown here are representative findings of an experiment from n = 3 independent experiments using different batches of Sertoli cells which yielded similar observations. Scale bar, 20 µm, which applies to other micrographs in this panel. B Representative IBs from n = 3 experiments with GAPDH served as the protein loading control (top panel). Composite data are shown in the lower panel, with each bar representing a mean ± SD of n = 3 experiments, illustrating that treatment of Sertoli cells with CdCl₂ at 3 µM for 24 h did not induce changes in Map1a expression even though its distribution across the cell cytoplasm was grossly perturbed as noted in A. However, p-p38-MAPK was induced by CdCl₂, and its up-regulation was blocked by doramapimod. Each sample had triplicate cultures in each experiment. ***, P < 0.005; ns, non-significantly different