Fig. 1.

Clozapine slows down the morphological and biochemical changes occurring during differentiation of SW872 cells.

(A) Representative images of ORO-stained LDs in differentiating SW872 cells grown for 10 days with or without CLZ (15 μM). Magnification 20X; scale bar represents 100 μm. (B) ORO quantification by spectrophotometric analysis at 510 nm. CTR samples received DMSO alone. Data are expressed in % respect to T0 condition. Results represent the means ± SEM calculated from at least three independent determinations, *p < 0.05, as compared to the CTR samples (Unpaired t-test). (C) Quantitative real time PCR of C/EBPβ expression after 3 days of differentiation (T3) with or without CLZ. The graph shows the normalized fold change compared to T0. GAPDH was used as housekeeping. (D) Western immunoblotting analysis of C/EBPβ. GAPDH was used as loading control. (E) Western immunoblotting analysis of PPARγ. Actin was used as loading control. T3 CTR received DMSO alone. Data are expressed in % respect to T0. Results represent the means ± SEM calculated from at least three independent determinations. **p < 0.01, as compared to T0; (**)p < 0.01, as compared to T3 (one-way ANOVA followed by Dunnett's test). (F–G) Immunocytochemical analysis of C/EBPβ and PPARγ expression at T3 with or without CLZ (Magnification 40X; scale bar represents 40 μm). Immunolocalization was visualized using diaminobenzidine (brown). Nuclei were counterstained with hematoxylin (blue grey). (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)