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. 2020 Nov 24;7:596366. doi: 10.3389/fmolb.2020.596366

FIGURE 5.

FIGURE 5

ExRNAs of Aa activate IL-6 in BV2 cells through the NF-κB signaling pathway. (A) Secreted IL-6 protein levels were upregulated at 16 h after treatment with intact OMVs. IL-6 secretion by OMV lysate was decreased by RNase-only treated OMV lysates in BV2 cells at 16 h. (B) qRT-PCR analysis revealed that the transcript levels of IL-6 activation by Aa OMV lysates were decreased by RNase-only pretreatment at 16 h. (C) NF-κB activation (phosopho-p65, upper panel) was significantly decreased by RNase-only-treated OMV lysates compared with DNase-treated OMV lysates at 16 h. Total NF-κB p65 (middle panel) and actin (bottom panel) levels were assessed for controls. BV2 cells were seeded onto 6-well plates (5 × 105 cells/well) and treated with Aa OMVs (approximately 4.5 × 108 particles/well in 2 ml of media) and OMV lysates (the same amount of proteins as in 4.5 × 108 OMV particles). Data are presented as mean ± SD from three independent experiments. The letters (a–c) indicate significant differences at p < 0.05.