GRA7-I-induced inflammasome activation was required for antimicrobial activity in MTB-infected macrophages.
2017-01-26T00:33:20Z (GMT) by
<p>(<b>A</b> and <b>B</b>) BMDMs from PKCα<sup>+/+</sup> and PKCα<sup>-/-</sup> mice (<b>A</b>) and BMDMs were transduced with lentivirus-shRNA-NS or lentivirus-shRNA-ASC for 2 days (<b>B</b>) infected with MTB (MOI = 1) for 4 h and then stimulated with rGRA7 (1, 5, 10 μg/ml) and its mutants for 18 h. IB analysis for IL-1β p17, IL-18 p18, or caspase-1 p10 in supernatants (SN), ASC, NLRP3, pro-IL-1β, pro-IL-18, or pro-caspase-1 in whole-cell lysates (WCL). Actin was used as a loading control. (<b>C</b> and <b>D</b>) Intracellular survival of MTB was assessed by CFU assay. BMDMs were infected with MTB for 4 h, followed by treatment with rGRA7, and then lysed to determine intracellular bacterial loads. The data are representative of five independent experiments with similar results (<b>A</b> and <b>B</b>). Data shown are the mean ± SD of five experiments (<b>C</b> and <b>D</b>). Significant differences (*<i>P</i> < 0.05; **<i>P</i> < 0.01; ***<i>P</i> < 0.001) compared with rVector. CFU, colony-forming units. ns, not significant.</p>