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. 2021 Jun 22;6(12):e144913. doi: 10.1172/jci.insight.144913

Figure 2. MMP2 binds and precipitates with TLR2 and TLR4.

Figure 2

(A and B) HEK293T cotransfection followed by Flag pull-down using magnetic beads. Mmp2-Flag was cotransfected with Tlr2-HA, Tlr4-Myc, or both plasmids (A). Mmp9-Flag cotransfection with Tlr-plasmids or Myd88-HA plasmid was also evaluated (B). (C) Reverse co-IP of Mmp2-Flag or Mmp9-Flag cotransfected with Tlr2-HA, Tlr4-Myc, or both plasmids. HA or Myc pull-down using anti-HA agarose or Anti–c-MYC agarose beads. (D) Schematic of the different MMP2 constructs used in the co-IP to identify the binding domain. (E and F) Different MMP2 domains were deleted or expressed alone and tested for co-IP with Tlr2-HA. Black line indicates where membrane ends/was cut. All transfections were performed using Lipofectamine 3000, and protein was extracted 20–24 hours after transfection. IP, immunoprecipitation; IB, immunoblotting; WCE, whole-cell extract; Δ, deletion.