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. 2023 Nov 20;11(11):e007736. doi: 10.1136/jitc-2023-007736

Figure 2.

Figure 2

EZH2 knockdown moderately increases expression of MHC class II. (A) EZH2, CIITA, and CD74 mRNA expression in MHC-II-proficient cell lines after transfection with EZH2 siRNA or to non-targeting control (NTC). Values were normalized to GAPDH mRNA and calculated as fold expression values relative NTC siRNA. The data shown represents combined technical replicates derived from three to four independent experiments. (B) after 24–48 hours IFN-γ stimulation, whole cell protein lysates were extracted from siRNA-transfected MHC-II-proficient cell lines probed with monoclonal antibodies specific for the indicated proteins. Lysates were normalized to GAPDH expression. Data represent one of two experimental replicates. (C) Flow cytometry analysis of MHC-II-proficient cell lines transfected with siEZH2 or NTC and stained with fluorophore-conjugated HLA-DR antibodies. The histograms shown in C represent one of at least three independent experiments. CIITA, class II major histocompatibility complex transactivator II; EZH2, enhancer of zeste 2 polycomb repressive complex 2 subunit; IFN, interferon; HLA-DR, human leukocyte antigen-DR; MHC, major histocompatibility complexes; mRNA, messenger RNA.