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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 3. Macrophages induce MITF expression through TNFα, which is required for melanoma growth.

Figure 3

A. Real time qPCR analysis for TNFα in melanoma cell lines and stromal cells: fibroblasts (HFF), keratinocytes (HACAT), THP1 and macrophages (M1, M2) compared to non-transformed melanocytes (NHM). B. TNFα production in conditioned media from undifferentiated and differentiated macrophages detected by ELISA. A Western blot for MITF and pIKBα of WM266-4 lysates following treatment with macrophage conditioned media for 24hrs is shown. C. qPCR gene expression analysis of MITF following treatment with macrophage conditioned media 24hrs with or without the addition of 5ng TNFα blocking antibody for 24hrs. D. Colony formation assay of WM266-4 cells treated with conditioned media from THP1 derived M1 or M2 macrophages or control medium for 3 weeks. E. Colony formation assay of WM266-4 cells treated with conditioned media from human monocyte derived M1 or M2 macrophages or control medium for 3 weeks. F. Growth of BRafV600E-4434 melanoma allografts in WT or LysMcre/TNFαF/F mice. G. CD68 histology of BRafV600E-4434 melanoma samples from WT and LysMcre/TNFαF/F mice. The relative CD68 immunofluorescence intensity (n=3 tumors) and TNFα expression (n=5 tumors) is shown.