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The analysis of cytoskeleton microfilaments was performed using FITC-phalloidin (Sigma-Aldrich). Fluoriescein-labeled phalloidin was used as described by Dejana et al. (1988). Samples dried and mounted with mounting medium (Calbiochem, La Jolla, CA) were visualized with an Olympus BX50 and photographed with a Leica DC500 digital camera. The morphological analysis was carried out on cell samples allowed to adhere to circular glass coverslips 13 mm in diameter (Electron Microscopy Sciences [EMS], Fort Washington, PA). HA were fixed in 2% glutaraldehyde in 0.1 GUCY1B3 M sodium-cacodylate (EMS) buffer, pH 7.2, for 1 hr at 4��C and then postfixed in 1% osmium tetroxide (EMS) for 1 hr at 4��C. After dehydration in graded ethanol and critical-point drying using CO2 (Emscope-CPD 750), the coverslips were coated with vacuum-evaporated gold (Emscope-SM 300) and observed with a Hitachi S4000 field emission scanning electron microscope. Each experiment was repeated three times in triplicate, and the means and standard deviations for each value were calculated. Statistical analysis of results was performed using paired Student's t-test with the statistical software OriginPro 8.5. Differences were considered significant at P?=?0.05. We preliminarily analyzed whether MeHg treatment has an impact on HA survival, evaluating its cytotoxic effects by MTT and LDH release assays. Figure 1A reports the results of cell viability, showing that 24 hr and 72 hr after MeHg treatment cell survival was significantly reduced by approximately 40% (P?Pexidartinib cost by astrocyte necrosis as demonstrated in Figure 1B, where the values corresponding to both MeHg treatments are completely comparable with those of untreated control cells (P?=?0.12). One of the main events of astrocyte reactivity is hypertrophy check details of astrocytic processes sustained by a filamentous network, composed of microfilaments (MF), microtubules (MT), and intermediate filaments (IF), consisting mostly of vimentin and GFAP (Bramanti et al., 2010a). In our experimental model, GFAP protein expression was markedly decreased after 24 hr (P?
