Fig. 1

ATGL over-expression is associated with increased proliferation rate in cervical cancer cell lines. (a) HeLa and (b) Me-180 cells were transfected with ATGL plasmid and after 24, 48 and 72 h the proliferation was assayed by Trypan blue direct cell counting procedure. Data are expressed as mean ± SD (n = 3; * p < 0.05 vs. CTRL). Bottom panels showed the Western blot analysis of the transfection efficiency. The high exposure (h.e.) allows to show ATGL expression in the CTRL sample. β-Actin was used as loading control. HeLa cells were transfected with ATGL plasmid for 48 h and (c) spectrophotometric determination of lipid droplets content after Oil Red-O staining was assessed. Oil Red-O incorporated in lipid droplets was eluted in isopropanol for 10 min and relative absorbance measured at 510 nm was normalized on total proteins. Data are expressed as mean ± SD (n = 3; * p < 0.05 vs. CTRL). (d) BrdU incorporation was quantified in HeLa cells by immunofluorescence, after 24 h of 10 μM BrdU treatment. Images reported are representative of three independent experiments. Nuclei were stained 10 min with 1 μg/mL Hoechst 33342. Bar graph refers to the percentage of BrdU-positive cells. Data are expressed as mean ± SD (n = 3; * p < 0.05 vs. CTRL). (e) Proliferation assayed by CCK-8 colorimetric assay. Data are expressed as mean ± SD (n = 3; * p < 0.05 vs. CTRL). (f) The levels of cell cycle regulator Cyclin D1 were evaluated by Western blot analysis. Band intensity is indicated below the corresponding band and expressed as fold-change relative to CTRL. The images are representative of three independent experiments that gave similar results. β-Actin and ATGL were used as loading and transfection control, respectively