Figure 3.

TP-472 inhibit melanoma growth in long-term survival assays, tumor growth in in vivo xenograft melanoma mouse model and invasion and migration in in vitro cell culture model. (A). Indicated melanoma cell lines were treated with the different concentrations of GSK343, JQ1, and TP-472 for 2 weeks. Cell survival was then measured using clonogenic assays. Representative images are shown. (B,C). A375 cells were treated with various concentrations of TP-472 and analyzed for ability to grow in an anchorage-independent manner in soft agar assays. Representative images of soft agar assays are shown. Scale bar, 500 μm (B). Bar diagram showing the relative colony size for each condition in panel B (C). (D). A375 cells were injected subcutaneously into the flanks of NSG mice (n = 6). The mice were treated with either vehicle or TP-472 (20 mg/Kg body weight) via intraperitoneal injection three times a week. The average tumor volume for each week is plotted. (E). Representative tumor images of NSG mice in vehicle and TP-472 treated mice at the end of the experiment. (F). A375 under indicated conditions were analyzed for the migration using wound-healing assay. Representative images showing the extent of migration in TP-472 treated cells relative DMSO treated cells are presented on the left and the quantification is presented as a bar diagram on the right. (G). A375 under indicated conditions were analyzed for invasive capacity using Matrigel invasion assay. Representative images of TP-472 treated invaded cells relative DMSO treated invaded cells are shown on the left and the quantification is presented as a bar diagram on the right. Data represent the mean ± standard error of three biological replicates. ** p < 0.01 and *** p < 0.001.