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. 2022 Oct 15;298(12):102596. doi: 10.1016/j.jbc.2022.102596

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© 2022 The Authors

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

E₉₁NP₃activates DCs through the TLR-2 pathway. The DCs were cultured with E₉₁NP₃ and controls (P₃, unstimulated DCs). After 5 to 6 h of stimulation, the cell lysate was prepared, and Western blotting was done. Blots depict the level of the expression of (A) p65 (NF-κB) and MyD88, (B) p38 and phospho-p38, and (C) STAT3 and phospho-STAT3 proteins. To check the gene level expression of different cytokines, qRT–PCR was performed using cDNA prepared from RNA isolated from the E₉₁NP₃-stimulated DCs and controls. D–G, bar graphs depict the relative expression of (D) il6, (E) il12, (F) tnfα, and (G) il1β genes. H–K, the DCs generated from WT (DC^WT) and TLR-2^−/− (DC^TLR-2−/−) C57BL/6 mice were cultured with FITC-labeled-E₉₁NP₃ for 4 h. Later, the cells were monitored for the (H) uptake of FITC-labeled-E₉₁NP₃ by the DC^WT and DC^TLR2^−/−; (I–L) expression of MHC II, CD80, CD86, and CD40 by the flow cytometer. The data (mean ± SD) shown are representative of two to three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. cDNA, complementary DNA; DC, dendritic cell; MHC, major histocompatibility complex; NP, nanoparticle; qRT–PCR, quantitative RT–PCR; TLR-2, toll-like receptor-2.

E₉₁NP₃activates DCs through the TLR-2 pathway. The DCs were cultured with E₉₁NP₃ and controls (P₃, unstimulated DCs). After 5 to 6 h of stimulation, the cell lysate was prepared, and Western blotting was done. Blots depict the level of the expression of (A) p65 (NF-κB) and MyD88, (B) p38 and phospho-p38, and (C) STAT3 and phospho-STAT3 proteins. To check the gene level expression of different cytokines, qRT–PCR was performed using cDNA prepared from RNA isolated from the E₉₁NP₃-stimulated DCs and controls. D–G, bar graphs depict the relative expression of (D) il6, (E) il12, (F) tnfα, and (G) il1β genes. H–K, the DCs generated from WT (DC^WT) and TLR-2^−/− (DC^TLR-2−/−) C57BL/6 mice were cultured with FITC-labeled-E₉₁NP₃ for 4 h. Later, the cells were monitored for the (H) uptake of FITC-labeled-E₉₁NP₃ by the DC^WT and DC^TLR2^−/−; (I–L) expression of MHC II, CD80, CD86, and CD40 by the flow cytometer. The data (mean ± SD) shown are representative of two to three independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. cDNA, complementary DNA; DC, dendritic cell; MHC, major histocompatibility complex; NP, nanoparticle; qRT–PCR, quantitative RT–PCR; TLR-2, toll-like receptor-2.