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. 2021 Nov 26;219(1):e20210789. doi: 10.1084/jem.20210789

Figure 2.

Figure 2.

AZD1775 induced ERVs transcription are responsible for IFN responses in ovarian cancer cells. (A) Western blot of basic expression of dsDNA and dsRNA sensor proteins in 11 ovarian cancer cell lines. Data represent three independent experiments. (B) Heatmaps of differentially expressed ERVs after AZD1775 for 48 h in ID8 cells. (C and D) Quantification of nine randomly selected ERVs by qPCR in ID8 and OV90 cells treated with DMSO or AZD1775 for 48 h (three independent experiments). (E) Cellular dsRNA was evaluated with anti-dsRNA (J2) immunofluorescence in ID8 cells with DMSO or AZD1775 for 72 h. RNase III was used as a negative control for dsRNA signal. Scale bars, 20 µm (three independent experiments). (F) Quantification of ISGs by qPCR in OV90 cells after AZD1775 for 48 h (three independent experiments). (G) MAVS protein polymerization detected by immunoblot using SDD-AGE (Materials and methods) in ID8 and OV90 cells treated with DMSO or AZD1775 for 72 h. Data represent three independent experiments. (H) Heatmap of relative expression of ISGs are shown. OVCAR4 cells were transfected with or without siRNA as indicated and treated with DMSO or AZD1775 for 48 h. Data are representative of three experiments. (I) Heatmap of differentially expressed genes after AZD1775 detected by NanoString immune panel in OVCAR4 cells with siRNA inhibition (n = 1) of the indicated dsDNA/dsRNA sensors. The qPCR data were normalized to β-actin. Data across panels represent mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001, as determined by unpaired t test (C, D, and F).