Fig 2. Blk is required for RNA or DNA virus-induced innate antiviral response in murine cells.
(A-C) Effects of Blk deficiency on SeV-, VSV- or EMCV-induced transcription of downstream genes and phosphorylation of TBK1 and IRF3. A20 cells were transduced with control (Con) or the indicated gRNA plasmids targeting Blk by the CRISPR/Cas9 method to establish the stable cell lines. Blk-deficient and control cells (2 × 105) were then uninfected or infected with viruses (MOI, 1) for the indicated times before qPCR (A) and immunoblot (B and C) analyses. (D and E) Effects of Blk deficiency on VSV- or HSV-1-induced transcription of downstream genes and phosphorylation of TBK1 and IRF3. pDCs derived from the spleens of Blk+/+ and Blk-/- mice (1 × 105) were infected with VSV or HSV-1 (MOI, 0.5) for 8 h before qPCR analysis (D) or infected with VSV (MOI, 0.5) for the indicated times before immunoblot analysis (E). (F and G) Effects of Blk deficiency on the replication of VSV. (F) pDCs derived from the spleens of Blk+/+ and Blk-/- mice (1 × 105) were infected with VSV-GFP (MOI, 0.2) for 8 h before flow cytometry analysis. (G) Statistical analysis in the form of a bar graph for the data in F. Graphs show mean ± SD (n = 2 technical replicates in A and D, n = 3 biological replicates in G) from one representative experiment. Data are representative of at least three independent experiments with similar results. *P < 0.05, **P < 0.01, ***P < 0.001 (unpaired, two-tailed Student’s t-test).