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. 2020 Oct 28;12(11):1221. doi: 10.3390/v12111221

Figure 1.

Figure 1

Caspase (CASP)8 activity is required for ∆M36-induced apoptosis. (A) Relative viability of bone marrow-derived macrophages (BMDM), bone marrow-derived dendritic cells (BMDC), murine embryonic fibroblasts (MEF), and SVEC4-10 cells infected for 24 h (h) with K181 or ∆M36 at MOI = 10. Relative viability was assessed with respect to parallelly maintained uninfected cells. All viability data are from 2 to 5 independent experiments. (B,C) Immunoblot (IB) for activated cleavage products of CASP8 (Cl-C8, 18 kDa) (B) and CASP3 (C) (Cl-C3; 22 and 19 kDa) in splenocytes (B) or splenic hematopoietic cells (HC; C) from mice infected with K181 or ∆M36 at indicated hours post-infection (hpi). IB are representatives of data obtained from 10 to 12 mice in 3 to 5 independent experiments for either B or C. (D) Relative viability of BMDM and BMDC cells at 24 hpi with K181 or ∆M36 in the absence or presence of zVAD-fmk (25 μM). The drug was added 1 hpi with virus and maintained till the end of the assay. E and (F) CASP8 activity on substrate sequence isoleucine-glutamine-threonine-aspartic acid (IETD) (E) and relative viability (B) of infected wild type (WT) BMDM from 6 through 20 hpi. (G) Relative viability of BMDM at 24 hpi when indicated genotypes were infected with K181 or ∆M36. Graphs show pooled data from 3 to 5 experiments for each condition. Lines above bars indicate the two groups compared for significance values using paired, parametric Student’s t-test where * is <0.05, ** is <0.01, and n.s. represents nonsignificant.