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. Author manuscript; available in PMC: 2020 Feb 1.
Published in final edited form as: Cell Rep. 2020 Jan 14;30(2):510–524.e6. doi: 10.1016/j.celrep.2019.12.036

Figure 5. FOXD3 Is Highly Expressed in Melanoma and Represses VISTA Expression.

Figure 5.

(A) RNA-seq data from TCGA was analyzed for FOXD3 expression across all of the cancer studies.

(B) Microarray data were reevaluated for FOXD3 regulation of immune checkpoint proteins.

(C) Expression of a V5-tagged FOXD3 was induced by treatment with doxycycline (100 ng/mL). After 48 h, doxycycline was refreshed and IFNγ (100 ng/mL) was added to select wells for an additional 48 h. Cells were lysed and protein expression was probed by western blot. Data are representative of 3 independent experiments.

(D) Expression of a V5-tagged LacZ control was induced by treatment with doxycycline for 96 h. Cells were lysed and protein expression was probed by western blot. Data are representative of 3 independent experiments.

(E) FOXD3 expression was induced by treatment with doxycycline for 96 h, and the cell surface levels of VISTA were analyzed by flow cytometry. Shown is the MFI. Data are representative of 3 independent experiments. *p < 0.05.

(F) A panel of lysates from various melanoma cell lines was probed for the expression of endogenous levels of VISTA and FOXD3 by western blot.

See also Figure S6.