Journal: Journal of Experimental & Clinical Cancer Research : CR
Article Title: The α-melanocyte stimulating hormone/peroxisome proliferator activated receptor-γ pathway down-regulates proliferation in melanoma cell lines
doi: 10.1186/s13046-017-0611-4
Figure Lengend Snippet: Analysis of PPARγ translocation into the nucleus and activity in response to 3 M3 exposure. ( a , b , c ) Immunofluorescence analysis of PPARγ localization in untreated cells ( A - C , G - I ) and in cells treated with 15 μM 3 M3 for 3 h ( D - F , J - L ). Immunolabeling with anti-PPARγ antibody and nuclear staining with DAPI. Scale bar: 20 μM. ( c ) Quantitative analysis of the PPARγ/DAPI colocalization signal in the nucleus. Results are express as fold increase of colocalization signal with respect to the values obtained in untreated cells and are reported as mean value ± SD (%) (* p < 0.01). ( d ) Transcriptional activity of PPARγ (fold change) by luciferase activity assay in B16-F10 and Mel 13. Cells were transfected with pGL3-(Jwt)3TKLuc reporter construct. After 24 h of transfection, cells were treated with 15 μM 3 M3. The measurement of luciferase activity was carried out 24 h after treatment. The variability of transfection was normalized with β -Gal activity. The results were expressed as fold change with respect to untreated cells. Data are mean values ± SD of three independent experiments performed in triplicate. * p < 0.01 (vs untreated cells)
Article Snippet: Quantitative analysis of the extent of PPARγ/DAPI colocalization was determined using Axiovision colocalization module (Zeiss) analyzing 100 cells for each condition randomly taken from 10 different microscopic fields.
Techniques: Translocation Assay, Activity Assay, Immunofluorescence, Immunolabeling, Staining, Luciferase, Transfection, Construct