Fig. 5

Combined treatment with g-PPT and Gefitinib decreases intracellular LD content by inhibiting SCD1 expression (a) The indicated NSCLC cell lines were exposed to concentrated depend g-PPT (100 nM, 1 μM, 10 μM, 20 μM) for 48 h and then harvested for Western blotting to detect the indicated signaling proteins. A representative blot is shown from three independent experiments. b Cell lines were seeded and exposed to vehicle (DMSO, NT), g-PPT (20 μM) for 48 h and then incubated in BODIPY for flow cytometer analysis. c Up panel, the indicated NSCLC cell lines were seeded and exposed to vehicle (DMSO, NT), g-PPT (20 μM), Gefitinib (2 μM) or g-PPT (20 μM) + Gefitinib (2 μM) for 48 h, and LD content was assessed by Nile red staining. Down panel, quantitation of lipid accumulation of indicated cell lines was evaluated by measuring Nile red fluorescence by flow cytometry; data are expressed as the percentage of level found in control cells treated only by the vehicle (HCC827GR DMSO) and are the means ± SD of at least 3 independent experiments. *p < 0.05, **p < 0.01, when compared to control cells. d Left panel, Edu staining was used to quantify the inhibition of cell proliferation. Cell lines were exposed to vehicle, g-PPT, Gefitinib or g-PPT + Gefitinib for 48 h and stained for Edu. Right panel, the proliferation rate was determined by counting the proliferating cells using ImageJ software. p values were determined by one-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001. e Left panel, the indicated cell lines were exposed to vehicle, g-PPT, Gefitinib or g-PPT + Gefitinib for 48 h, and the content of LDs and expression p-EGFR1068 were assessed by IF. Right panel, quantitation of p-EGFR expression of indicated cell lines. f Left panel, the given cell lines after exposed for 48 h and assessed the expression of lipid droplets and SCD1 by immunofluorescence. Right panel, quantitation of SCD1 expression of indicated cell lines. p value calculated as above.