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Figure 3 | Molecular Cancer

Figure 3

From: Timp1 interacts with beta-1 integrin and CD63 along melanoma genesis and confers anoikis resistance by activating PI3-K signaling pathway independently of Akt phosphorylation

Figure 3

The presence of Timp1 in the 4C11+ conditioned medium confers anoikis resistance to melanocytes. A. The MaGFP and MaT1S cell lines (2x102) were incubated at 37°C on plates of 60 mm2 adhered to the substrate for 7 days. After this period, the formation of clones was visualized by adding the dye toluidine blue in 1% borax. B. The melan-a melanocytes and 4C11+ melanoma cell lines were maintained in culture in RPMI pH 6.9 0.5% FCS in the presence of PMA for 48 hours. After this period, the medium conditioned by these cells was collected. Melan-a melanocytes were grown in suspension for 96 hours in three different conditions: a) with melan-a conditioned medium, b) with 4C11+ conditioned medium, or c) with 4C11+ conditioned medium in the presence of Timp1-neutralizing antibody. After 96 hours, the formation of clones by clonogenic assay was analyzed. C. After 96 hours of anchorage blockage, all cells were collected and viable cells were determined by MTT assay. maGFP: murine non-tumorigenic melanocyte lineage transfected with fluorescent protein GFP; maT1S: murine non-tumorigenic melanocyte overexpressing Timp1; ma: non-tumorigenic melan-a melanocytes; 4C11+: metastatic melanoma cells. **p < 0.01, ***p < 0.001.

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Molecular Cancer

ISSN: 1476-4598