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. 2017 Mar 21;31(7):2881–2892. doi: 10.1096/fj.201601238R

Figure 4.

Figure 4.

TLR4-NF-κB p65/p50 pathway contributes to reduction of Nrp1 by LPS. A) Immunoblot analysis and quantification of Nrp1 in RAW264.7 cells transfected with scramble or TLR4 siRNA for 48 h and treated with 100 ng/ml LPS for 12 h. B) Immunoblot analysis and quantification of Nrp1 in RAW264.7 cells stimulated or unstimulated with LPS (100 ng/ml) for 12 h with or without incubation of NF-κB activation inhibitor (10 μM). Immunoblot analysis of Nrp1 in RAW264.7. β-Actin serves as loading control. C) Quantitative RT-PCR analysis of Nrp1 mRNA in RAW264.7 cells treated with LPS (100 ng/ml) alone or LPS (100 ng/ml) plus NF-κB activation inhibitor (10 μM) for indicated time. D–F) Quantitative RT-PCR analysis of Nrp1 mRNA in RAW264.7 cells transfected with scramble or NF-κB p50 (D), p65 (E), or p50/p65 (F) siRNA for 48 h and then treated with 100 ng/ml LPS for 12 h. Data are expressed as means ± sem. *P < 0.05 vs. control or indicated.