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. 2022 Mar 21;9:832713. doi: 10.3389/fmed.2022.832713

Figure 3.

Figure 3

The PEP-NASP significantly inhibits tumor necrosis factor receptor-associated factor 6 (TRAF6) polyubiquitination in LPS-stimulated RAW 264.7 cells. (A) Cells were pretreated with or without the PEP-NASP or NASP (0.4 μM) for 1 h before the addition of LPS (1 μg/mL) for 30 min. Cell lysates were immunoprecipitated with an anti-TRAF6 followed by immunoblotting (IB) with an antibody against Ub or TRAF6. (B) Immunoprecipitation (IP) of endogenous TRAF6 (with anti-TRAF6) from RAW 264.7 cells transduced with the PEP-NASP or NASP, followed by IB with an antibody against His, NASP, or TRAF6. Total cell lysate (TCL) IB was done with anti-NASP and anti-His. (C) Cells were transduced with the PEP-NASP or NASP, stimulated with LPS, and assessed by IB with an antibody against His, NASP, or TRAF6 after IP with anti-TRAF6 or by IB with anti-NASP and anti-His in TCL. Data represent a minimum of three independent experiments.