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. 2021 Feb 27;10(3):362. doi: 10.3390/antiox10030362

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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IMP interferes with LPS binding to LBP, CD14, and MD-2, in TLR4 co-receptor complex in RAW264.7 cells. RAW264.7 cells were pretreated with IMP for 1 h and then stimulated without or with 100 ng/mL of LPS for 24 h incubation. (a) Cells were pretreated with various IMP concentrations (0, 10, 20, 40, or 80 μM) or Dexa (2 μM) for 1 h and were stimulated with LPS for 24 h. The protein expressions of LBP, CD14, MD-2, TLR4, and MyD88 were measured by Western blotting (n = 3). (b) Cells were pretreated with 80 μM IMP or 2 μM Dexa for 1 h and were stimulated with LPS for 24 h. We also measured LBP by immunofluorescence staining. For LBP (green) in cells, nuclei were labeled with DAPI (blue). Alternatively, it was stained for CD14 (green) (c), or cells were stained for MD-2 (green) (d) and TLR4 (green) (e). Scale bar = 10 μm. The detailed experimental preforms are described in Materials and Methods (n = 3). Results (data) are presented as means ± S.E.M. ^# p < 0.05 compared with the control group, * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the LPS-alone group.

IMP interferes with LPS binding to LBP, CD14, and MD-2, in TLR4 co-receptor complex in RAW264.7 cells. RAW264.7 cells were pretreated with IMP for 1 h and then stimulated without or with 100 ng/mL of LPS for 24 h incubation. (a) Cells were pretreated with various IMP concentrations (0, 10, 20, 40, or 80 μM) or Dexa (2 μM) for 1 h and were stimulated with LPS for 24 h. The protein expressions of LBP, CD14, MD-2, TLR4, and MyD88 were measured by Western blotting (n = 3). (b) Cells were pretreated with 80 μM IMP or 2 μM Dexa for 1 h and were stimulated with LPS for 24 h. We also measured LBP by immunofluorescence staining. For LBP (green) in cells, nuclei were labeled with DAPI (blue). Alternatively, it was stained for CD14 (green) (c), or cells were stained for MD-2 (green) (d) and TLR4 (green) (e). Scale bar = 10 μm. The detailed experimental preforms are described in Materials and Methods (n = 3). Results (data) are presented as means ± S.E.M. ^# p < 0.05 compared with the control group, * p < 0.05, ** p < 0.01, and *** p < 0.001 compared with the LPS-alone group.