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IBD_23006_sm_SuppFig1.tif2170KSupporting Information Figure 1. Construction of mouse IL-10 expression vector pSC1m. S. cerevisiae codon optimized sequences for alpha-mating prepro region and mature mouse IL-10 were amplified from Geneart vector pGA12. To the amplicon's 3′ end, S. cerevisiae ADH I terminator and G418 resistance marker (conferring Kanamycin resistance) amplified from plasmid pKT127 were tagged by fusion PCR technique. The fusion PCR product was subsequently cloned into SpeI/BamHI digested yeast high copy plasmid p426GPD under control of the GPD promoter resulting in mouse IL-10 expression vector pSC1m. Kan, bla - genes conferring kanamycin and ampicillin resistance, respectively; aMF - alpha-mating factor prepro sequence; mIL-10 - mouse interleukin-10; ADH1 - yeast ADH I terminator sequence.
IBD_23006_sm_SuppFig2.tif252KSupporting Information Figure 2. Activation of STAT3 phosphorylation by S. boulardii-derived recombinant mouse IL-10. STAT3 phosphorylation (STAT3-pY705) was assessed by immunoblot analysis on protein extracts of J774.1 macrophage cells treated with supernatant from pSC1m transformed S. boulardii containing recombinant mouse IL-10 at 150 and 300 ng/ml, respectively. Total STAT3 was used as loading control to ensure equal amounts of protein in all lanes. Supernatant of p426GPD-Kana transformed S. boulardii and commercial mouse IL-10 served as controls. One experiment representative of two is shown. SN - supernatant; mIL-10 - mouse interleukin-10.

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