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. 2017 Mar 7;7:44252. doi: 10.1038/srep44252

Figure 3. Neoagarotetraose decreased protein and mRNA levels of pro-inflammatory cytokines in LPS-stimulated RAW264.7 cells.

Figure 3

(A,B) After the pre-treatment of cells with different concentrations of neoagarotetraose (62.5, 125, 250, 500 μg/ml) for 2 h, the LPS (100 ng) was added to cells and incubated for 16 h. Then the protein levels of pro-inflammatory cytokines TNF-α (A) and IL-6 (B) in cell culture media were measured using ELISA kits in a microplate reader, respectively. Values are the means ± SD (n = 3). Significance: ##P < 0.01 vs. normal control; *P < 0.05, **P < 0.01 vs. LPS treated control. (C,D) The total mRNAs of RAW264.7 cells were collected after treatment with neoagarotetraose (62.5, 125, 250, 500 μg/ml) for 2 h and 100 ng LPS for 16 h. Then the mRNA levels of TNF-α gene (C) and IL-6 gene (D) were detected by quantitative RT-PCR assay, respectively. The relative amounts of TNF-α and IL-6 mRNAs were determined using the comparative (2−ΔΔCT) method. The mRNA levels for non-drug treated cells (Control) were assigned values of 1. Values are means ± S.D. (n = 3). Significance: #P < 0.05, ##P < 0.01 vs. normal control; *P < 0.05, **P < 0.01 vs. LPS treated control.