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[Preprint]. 2023 Jun 8:2023.06.06.543919. [Version 1] doi: 10.1101/2023.06.06.543919

Figure 1: EZH2 inhibition allows up-regulation of MHC Class I and Class II in 2D human LSCC cell lines.

Figure 1:

A) Schematic for proposed mechanism: Inhibition of EZH2 methyltransferase activity by the drugs GSK126 or EPZ6438 will lead to de-repression of antigen presentation genes that can then be more effectively activated by interferon gamma. B) RT-qPCR in the indicated four human lung cancer cell lines treated for 7 days with vehicle or EZH2 inhibition with IFNγ added on day 5 for the genes B2M, HLA-A, CIITA and HLA-DRA, mean +/− SEM is graphed, n = 4 individual cultures, *indicates p<0.04, **p<0.008 ***p<0.0009, ****p<0.0001 by one-way ANOVA with pairwise comparisons and Holm-šídák’s post hoc test. C) Flow cytometry analysis of indicated four human lung cancer cell lines treated for 7 days with vehicle or EZH2 inhibition with IFNγ added on day 5 for the cell surface proteins HLA-A,B,C and HLA-DR, mean +/− SEM is graphed, n = 4 individual cultures, * indicates p<0.04, ** p<0.006, ***p<0.0009, ****p<0.0001 by one-way ANOVA with pairwise comparisons and Holm-šídák’s post hoc test. Representative histograms from HCC15 cell lines are shown, G=GSK126, E=EPZ6438, I=IFNγ, I+G= IFNγ+GSK126 and I+E= IFNγ+EPZ6438. D) Western blotting of A549 and HCC15 cell lines treated for 7 days with vehicle or EZH2 inhibition with IFN³ added on day 5 for the proteins B2M, HLA-DR,DQ,DP, EZH2, H3K27me3 and total histone H3. Data are representative of two individual cultures. See also Supplementary Figure 1.