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. 2019 Apr 2;20(7):1629. doi: 10.3390/ijms20071629

Figure 3.

Figure 3

Treatment with fatty acids (PAOA) increases neutral fat content in 3D multilineage spheroids. (A) Intracellular neutral lipid content visualized by ORO staining in sections (8 μm) of 3D spheroids HepG2/LX-2 ratio 24:1 treated, after 48 h from the seeding with BSA 1%, a mix of palmitic acid and oleic acid (PAOA) 500 μM (1:2), TGF-β 10 ng/mL or PDGF 10 ng/mL for 48 h. Cell nuclei were stained with hematoxylin. Objective: 40×. (B) Quantification of intracellular ORO-stained area quantified by BioPix software (n = 5). (C) Intracellular lipid content measured by AdipoRed assay kit (Lonza). Bars represent mean ± SD (n = 6). P-value was calculated by Mann-Whitney non-parametric test, (** p < 0.005 vs. BSA 1%). BSA: bovine serum albumin; PAOA: palmitic acid/oleic acid; TGF-β: Transforming growth factor β; PDGF: Platelet-derived growth factor, ORO: oil red O staining.