KDM5 histone demethylases repress immune response via suppression of STING
Fig 2
Inhibition of KDM5 demethylases primes the innate antiviral immune response.
(A) Representative images of MCF7 cells 24 hours after infection with VSV-GFP viruses at MOI 0.5. Scale bar, 100 μm. (B) Flow cytometry plot (left panel, 24 hours) and quantification of GFP-positive cells (right panel) after infection with VSV-GFP virus for the indicated time at MOI 0.5. (C) qPCR analysis of DNA copy number of vaccinia virus in MCF7 cells at the indicated time after infection at MOI 0.25. (D) Representative crystal violet staining images (left panel) and quantification of relative intensity (right panel) of MCF7 cells 3 days after infection with vaccinia virus at MOI 0.5. (E) qPCR analysis of DNA copy number of vaccinia virus in growth media from the cells shown in panel D. (F) Quantification of flow cytometry analysis for the percentage of GFP-positive cells in control or KDM5B and KDM5C double knockout cells 12 hours after infection with VSV-GFP virus at MOI 0.5. (G) Representative crystal violet staining images (left panel) and quantification of relative intensity (right panel) of the indicated MCF7 knockout cells 3 days after infection with vaccinia virus at MOI 0.25. (H) qPCR analysis of DNA copy number of vaccinia virus in growth media from the cells shown in panel G. Cells were pretreated with DMSO or 1 μM KDM5-C70 for 5 days, followed by no treatment for 1 day, before viral infection in panel A–E. Representative data from triplicate experiments are shown in panel C, E, F, and H. Two or 3 biological replicates are shown in panel B, D, and G. Error bar denotes SEM. #p < 0.01. The numerical values used to generate graphs in B–H are available in S1 Data. MOI, multiplicity of infection; qPCR, quantitative PCR; VSV-GFP, vesicular stomatitis virus carrying a green fluorescent protein reporter.