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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: The α-melanocyte stimulating hormone/peroxisome proliferator activated receptor-γ pathway down-regulates proliferation in melanoma cell lines

Fig. 6

The PPARγ acts as an effector element in determining the reduction of cell number induced by the activation of PI(4,5)P2/PLC pathway in B16-F10 and Mel 13 melanoma cell lines. a Analysis of cell number performed on B16-F10 and Mel 13, after treatment for 24 and 48 h with 15 μM 3 m3 or 10−7 M αMSH in the presence or absence of 3 μM GW9662. In the combined treatment αMSH/GW9662 or 3 m3/GW9662, cells underwent a pre-treatment with GW9662 for 1 h. Cell number was expressed as a percent variation in comparison to the value of untreated control cells. Data are mean values ± SD of three independent experiments performed in triplicate. *p < 0.01 and **p < 0.001 (vs untreated cells); $ p < 0.01 (vs 3 m3-treated cells); § p < 0.01 (vs αMSH-treated cells). b Cell-cycle distribution evaluated by flow cytometric analysis on B16-F10 and Mel 13, after treatment for 24 h with 15 μM 3 m3 or 10−7 M αMSH in the presence or absence of 3 μM GW9662. In the combined treatment αMSH/GW9662 or 3 M3/GW9662, cells underwent a pre-treatment with GW9662 for 1 h. The bar graph shows the distribution of cells among the different phases of the cell cycle. Data are mean values ± SD of three independent experiments performed in duplicate. *p < 0.01 (vs untreated cells); $ p < 0.01 (vs 3 m3-treated cells); § p < 0.01 (vs αMSH-treated cells). c Western blot analysis of p27 (4 h), p21 (48 h), cyclin D1 and cyclin E (6 h) protein expression on cell lysate of B16-F10 and Mel 13, treated with 10−7 M αMSH in the presence or absence of 3 μM GW9662. In the combined treatment αMSH/GW9662, cells underwent a pre-treatment with GW9662 for 1 h. GAPDH or HSP70 were used as an equal loading control. Results refer to three independent experiments. Representative blots are shown. Densitometric scanning of band intensities was performed to quantify the change of protein expression (control value taken as one fold in each case). * p < 0,01 (vs untreated cells), $ p < 0,01 (vs αMSH-treated cells)

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