Figure 1

PPARγ inhibition results in ERBB2-positive cancer cell death and increased levels of fats. (a) Cell counts of BT474, MDA-MB-361, MCF-7, and human mammary epithelial cell (HMEC) cells treated with vehicle, 5, 10, and 20 μmol/l of the peroxisome proliferators-activated receptor (PPAR)γ antagonist GW9662 for 24, 48, and 72 hours. Results are presented as percentage of control (vehicle). Error bars indicate the standard deviation from three individual experiments. ***P < 0.003, **P < 0.05. (b) Quantification (nmol/100,000 cells) of total cellular fats from BT474 and MCF-7 whole cell extracts by mass spectrometry after treatment with vehicle or 10 μmol/l of GW9662. Error bars indicate the standard deviation from two experiments. **P = 0.01, *P = 0.3). (c) Quantification of neutral fat stores of BT474 and MCF-7 cells treated with vehicle or 10 μmol/l of GW9662 by using the BODIPY 493/503 lipid probe staining. Error bars indicate the standard deviation from three individual experiments. **P = 0.05, *P = 0.1. (d) RT-PCR of BT474 cells treated with vehicle, 10, or 20 μmol/l of GW9662. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as control. (e) Quantification (in nmol/100,000 cells) by mass spectrometry of palmitic, palmitoleic, stearic, and oleic fatty acids from BT474 and MCF-7 whole cell extracts after treatment with vehicle or 10 μmol/l of GW9662. Error bars indicate the standard deviation from two experiments. **P < 0.05. aP2, fatty acid binding protein 4 (FABP4); ATGL, adipose tryglyceride lipase; CPT, carnitine palmitoyltransferase; HSL, hormone sensitive lipase.