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. 2020 Apr 20;11:1897. doi: 10.1038/s41467-020-15726-7

Fig. 5. TGFß promotes HLA-ABC downregulation at baseline and in response to IFNγ.

Fig. 5

a IFNγ-mediated induction (IFNγ-treated/vehicle-treated control) of cell surface HLA-ABC in MITFhigh/AXLlow or /MITFlow/AXLhigh short-term PD-1 PROG melanoma cell lines. Each dot represents one cell line and HLA-ABC induction was measured by flow cytometry 24 h after treating cultures with vehicle control or 1000 U/ml IFNγ. Box plots show the median and interquartile ranges, and data were compared using Mann-Whitney test. b Cell surface expression (median fluorescence intensity; MFI) of HLA-ABC in WMD-084, SCC14-0257 and SMU17-0132 melanoma cells treated with vehicle (Control), 1000 U/ml IFNγ and/or 10 ng/ml TGFß for 72 h. Data (mean ± s.d.) were compared using one-way ANOVA with the Geisser-Greenhouse correction. c Expression of de-differentiation markers AXL, N-cadherin and SNAIL in WMD-084, SCC14-0257 and SMU17-0132 melanoma cells treated with vehicle (Control), 1000 U/ml IFNγ- and/or 10 ng/ml TGFß for 72 h. d IFNγ production after co-culture of TGFß pre-treated (10 ng/ml for 72 h) melanoma cells with the patient-matched tumor-infiltrating lymphocytes expanded from the same tumor biopsy. IFNγ was measured by flow cytometry. Data (mean ± s.d.) show relative IFNγ expression in T cells (TGFß pre-treated/BSA pre-treated) after background subtraction (spontaneous IFNγ production on immune cell-only cultures). Paired BSA-treated vs TGFß-treated data were compared using paired t-test, **p < 0.01, *p < 0.05.