Figure 2.

GNP treatment increased neutral lipid contents in activated cancer-associated fibroblasts (CAFs). (A-B). CAFs (CAF19, PTAF2 and PTAF3 were plated at a density of 10⁴ cells/well in a 12 well-plate with cover glass) followed treatment with GNPs (20 ug/ml) or remained non-treated (NT) in SCM or SFM condition. After 48h, neutral lipids were stained with BODIPY (green, λex./em.=493/503 nm) for 30 mins and were visualized under fluorescence microscope. Representative images of cells were shown here from three independent experiments. Scale bar 20 μm. (C-D). Quantification of lipid contents in CAFs cultured in SCM and SFM using Oil Red O. CAFs (2×10⁴ cells/well in 6-well plate) were prepared and were treated with GNPs or remained NT. Post 48h, lipids were stained with Oil Red O dye and that dye was extracted by using 100% isopropanol. In separate experiment, cells were counted by using hemocytometer. The optical density (OD) value of Oil Red O dye was measured at 500 nm and was expressed as OD/10⁴ cells. Statistical significance was determined by student’s unpaired t-test(NT vs GNPs, n=3, mean±s.d., *p<0.05, **p<0.01 and ****p<0.0001). (E) Images after Oil red o staining CAFs were prepared, stained with oil red o dye and were then visualized by Nikon Eclipse Ni microscope. Scale bar 20 μm.