Figure 9.

CEN induces nonprotective autophagy in SAOs cells. SAOs cells were treated with NE and CEN (200 μg/ml, w/v) in the presence of a si-RNA against BECN1 (si-BECN1) or its corresponding control si-RNA (si-FITC, fluorescein conjugate) for 72 h. CyQuant assay was performed at the end of incubation as described in Materials and Methods. (a) Bar graphs represent the mean ± SD; symbols indicate significance: ^∗∗∗p < 0.001, ^∗∗p < 0.01, and ^∗p < 0.05 versus NE and ⁺⁺⁺p < 0.001 and ⁺p < 0.05 versus untreated cells; no significant differences were detected between si-BECN1 versus si-FITC. (b) Representative micrographs (microscope Axiovert 200 Zeiss; FITC fluorescence filter, 200x magnification) of cell nuclei stained with CyQuant fluorescent dye of SAOs untreated (A), (D), (G), after 72 h treatment with NE (B), (E), (H) and CEN (200 mg/ml, w/v) (C), (F), (I) (legend on top). The treatments with si-BECN1 and si-FITC described referred to (G), (H), (I) and (D), (E), (F), respectively (legend on the left).