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. 2021 Feb 26;9(3):494. doi: 10.3390/microorganisms9030494

Figure 2.

Figure 2

Determination of the amino acid residues of NSP1 critical for caspase-1 inhibition. (A and B) HEK 293T cells were co-transfected with vectors expressing caspase-1 and pro-IL-1β plus either pEF1α-DEST or pEF1α-DEST expressing c-Myc-tagged NSP1 wild type (WT) or NSP1 containing the mutations R124A/K125A, K164A/H165A, and both R124A/K125A and K164A/H165A (quadruple mutant (QM)) and a deletion of amino acids 160–173 (∆ 160–173). After 24 h, (A) the concentrations of mature IL-1β secreted in culture supernatants were measured by ELISA, and (B) levels of caspase-1 and NSP1 (c-Myc) proteins and the cleavage of caspase-1 into its p20 subunit were determined by Western blotting. ELISA data shown are representative of three independent experiments. * p < 0.001 by unpaired two-tailed Student’s t-tests.