fig3

Figure 3. Caco-2 polarized monolayers released TGs in response to FFAs and more so in the co-presence of LPS. Fat accumulation in Caco-2 polarized monolayers on top of the co-culture system was evaluated by ORO staining (200× magnification) and quantified by ImageJ software in 10 random non-overlapping micrographs for each experimental condition, by calculating the percentage of pixels above the threshold value with respect to the total pixels per area. At least three independent experiments were conducted (A, B). Intracellular TG content was assessed in lysates from Caco-2 polarized monolayers (C). Assessment of APOB (D), TG-rich lipoproteins (E), TGs (F) and FFAs (G) secreted were measured in concentrated culture media from the basolateral compartment and normalized for the total protein content. For violin plots, data were expressed as median concentration (thick dashed lines) and interquartile range (dotted lines). At least three independent experiments were conducted and pooled together. ***P < 0.0001, *P < 0.01 according to one-way ANOVA. TGs: Triglycerides; FFAs: free fatty acids; LPS: lipopolysaccharides; ORO: oil red O; APOB: apolipoprotein B; ANOVA: one-way analysis of variance.