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. 2016 Sep 8;5:e16432. doi: 10.7554/eLife.16432

Figure 3. IFI6 is necessary for NRAS-mutant melanoma tumor growth.

(A) NRAS-mutant melanoma cell lines expressing either non-specific (NS) or IFI6 shRNA were analyzed for anchorage-independent growth using the soft agar assay. Representative images for the indicated melanoma cell lines expressing NS or IFI6 shRNA are shown. (B) Relative colony size in the soft agar assay presented in panel A are plotted for the indicated melanoma cell lines expressing either NS or IFI6 shRNA. (C) The indicated NRAS-mutant melanoma cell lines expressing either NS or IFI6 shRNA were injected subcutaneously into the flank of athymic nude mice. Average tumor volumes (n = 5) at the indicated time points are shown. (D) YUVON cells expressing NS or IFI6 shRNA were analyzed for IFI6 mRNA expression by RT-qPCR. Relative IFI6 mRNA expression (%) in YUVON cells expressing IFI6 shRNA in comparison to NS shRNA-expressing cells is shown. (E) YUVON cells expressing NS or IFI6 shRNA were analyzed for their ability to grow in soft agar. Representative wells of YUVON cells expressing the indicated shRNA and microscopic images are shown. (F) YUVON cells expressing NS or IFI6 shRNA were injected subcutaneously in athymic nude mice. Average tumor volumes (n = 5) for the indicated time points are shown. In all panels, data are presented as mean ± SEM, and *p<0.05 and **p<0.005.

DOI: http://dx.doi.org/10.7554/eLife.16432.012

Figure 3.

Figure 3—figure supplement 1. IFI6 is necessary for NRAS-mutant melanoma cell growth.

Figure 3—figure supplement 1.

Representative soft agar well images of NRAS-mutant melanoma cell lines expressing non-specific (NS) or IFI6 shRNAs.
DOI: http://dx.doi.org/10.7554/eLife.16432.013
Figure 3—figure supplement 2. Ectopic expression of shRNA-resistant IFI6 ORF rescues tumor growth in YUGASP cells expressing IFI6 shRNA.

Figure 3—figure supplement 2.

(Top panel) Anchorage-independent growth in soft agar was evaluated using YUGASP cells expressing non-specific (NS) shRNA or IFI6 shRNA with or without the shRNA-resistant IFI6 open reading frame (ORF). Representative images under the indicated conditions are shown. (Middle panel) YUGASP cells expressing the indicated constructs were analyzed for relative protein levels of IFI6 or ACTINB by immunoblotting. (Bottom panel) YUGASP cells expressing the indicated constructs were injected subcutaneously into athymic nude mice. Average tumor volumes (n = 8) under at indicated times are shown. Data are presented as mean ± SEM; **p<0.005.
DOI: http://dx.doi.org/10.7554/eLife.16432.014