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. Author manuscript; available in PMC: 2015 Apr 1.
Published in final edited form as: Cancer Discov. 2014 Oct;4(10):1214–1229. doi: 10.1158/2159-8290.CD-13-1007

Figure 1. TNFα is an important survival and growth signal for melanoma.

Figure 1

A. Kaplan-Meier plot showing melanoma-free survival (%) of tamoxifen-treated BrafV600E;Tyr::CreERT2 (BRAFV600E) and BrafV600E;Tnfα−/−;Tyr::CreERT2 (BRAFV600E/TNFα−/−) mice, and control mice (Ethanol-treated BrafV600E;Tnfα−/−;Tyr::CreERT2 mice and tamoxifen-treated Tyr::CreERT2 mice). p< 0.0001; Log-rank (Mantel Cox) Test. B. Growth of BRafV600E-4434 melanoma allografts in WT and TNFα−/− mice. C. in vitro growth assay of BRafV600E-4434 melanoma cells treated with BSA or 50ng TNFα once every 3 days. D. Anoikis assay of BRafV600E-4434 melanoma cells for dead cells detected by trypan blue staining. Cells were cultured under non-adherent conditions for 72hrs and treated with BSA or 50ng TNFα. A Western blot for MITF, pIKBα, cleaved caspase3 and ERK2 is shown. E. Western blot of the indicated cell lines for MITF and pIKBα and ERK2 after 24hrs treatment with 50ng TNFα. F. Anoikis assay for untreated or TNFα treated 4434, A375 and 4434- and A375-MITF overexpressing cells. G. Western blot for MITF, pIKBα, cleaved caspase3 and ERK2 of detached A375 and A375-MITF cells treated for 48hrs with 50ng TNFα. H. Anoikis assay for untreated or TNFα stimulated A375 cells transfected with control or MITF specific siRNAs. A Western blot for MITF, pIKBα, cleaved caspase3 and ERK2 is shown.