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. 2021 Jul 23;12:685984. doi: 10.3389/fimmu.2021.685984

Figure 6.

Figure 6

AC-derived Hexa act as a key ingredient in promoting M2 activation of macrophages. (A) Diagram of isolation of 10~20 kDa-AC L4 exofree using ultrafiltration tubes, and the efficiency was confirmed using western blot. (B) Heatmap showed the relative frequencies of amino acids between the asparagine (Asn) position of N-linked glycoproteins in AC L4 exofree. Enrichment level of the amino acid at indicate position is shown in different colors. Red means high enrichment level, and green means low enrichment level. (C) Venn diagram of AC L4 exosome, AC L4 exofree proteins identified using mass spectrum analysis. (D) Diagram of Bi-layer PPI network analysis using AC L4 exofree proteins and RNA-seq data of BMDMs. (E) GO enrichment analysis (Biological process) of exofree-induced TCGs of BMDMs ( Supplementary Figure 5B ). (F) A protein-gene network was constructed for worm proteins with molecular weight less than 50kDa and TCGs of exofree-, IL-4-, and exofree+IL-4-treated BMDMs ( Supplementary Figure 5A ), based on the LC-MS/MS and RNA-seq data. (G) Phylogene tree of Hexa protein in 17 species. (H, J) qPCR analysis of mRNA level of Arg1 (H) and Chi3l3 (J) were measured in the presence of PBS, Hexa 5 ng/ml, Hexa 50 ng/ml, Hexa 500 ng/ml, IL-13 10 ng/ml, IL-13+Hexa 5 ng/ml, IL-13+Hexa 50 ng/ml, IL-13+Hexa 500 ng/ml for 24 h. (I, K) qPCR analysis of mRNA level of Arg1 (I) and Chi3l3 (K) were measured in the presence of PBS, Hexa 5 ng/ml, Hexa 50 ng/ml, Hexa 500 ng/ml, IL-4 10 ng/ml, IL-4+Hexa 5 ng/ml, IL-4+Hexa 50 ng/ml, IL-4+Hexa 500 ng/ml for 24 h. Data information: *P<0.05, ***P<0.001, ****P<0.0001.