NLRP3 Activation Was Regulated by DNA Methylation Modification during Mycobacterium tuberculosis Infection

Figure 2

Construction and identification of NLRP3 promoter. (a) Two potential promoter regions of NLRP3 were amplified from genome and identified by double restriction enzyme digestion. 1 and 2 were PCR products from human genomic DNA. P1 and P2 were pNLPR3-P1 and pNLPR3-P2, respectively. P1E and P2E exhibited the enzyme digested products of pNLPR3-P1 and pNLPR3-P2 digested by Xho I and Mlu I. M1 and M2 were 250 bp and DL2000 DNA marker, respectively. (b) The activities of two potential NLRP3 promoters were identified by Dual-Luciferase Reporter Assay. HEK293T cells were plated in 24-well cell culture plate and cotransfected with firefly luciferase report plasmids pNLRP3-P1/P2 and Renilla luciferase report plasmid pRL. The relative promoter activity was measured 48 h after transfection as in Materials and Methods. Firefly luciferase activity was normalized to Renilla luciferase activity, and the promoter activity was expressed as the mean ± SD of at least triplicate wells (; NS: no significant difference).

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