P stat3 s727 pe

Spleen tissues were isolated from mice in both the group at 14 weeks after curdlan injection. To examine the populations of Th17 and Treg cells, the tissues were stained with Abs against CD4–fluorescein isothiocyanate (FITC), IL-17–phycoerythrin (PE), CD25–allophycocyanin, and FOXP3–PE (all from eBioscience, San Diego, CA, USA). To determine the population of cells expressing STAT, the tissues were stained with Abs against CD4–FITC, p-STAT3–Y705–PE, and p-STAT3–S727–PE (eBioscience, San Diego, CA, USA). The stained tissue sections were analyzed using a confocal microscope (LSM 510 Meta; Carl Zeiss, Oberkochen, Germany). We counted cell number using Pannoramic MIDI and Pannoramic viewer (3D HISTECH Ltd, Hungary).

Spleens were obtained from mice in both groups at 10 weeks after curdlan injection. To assess the differentiation of Th17 cells and Tregs, the spleen tissues were stained with Abs against CD4–fluorescein isothiocyanate (FITC), IL-17–phycoerythrin (PE), CD25–allophycocyanin, and forkhead box P3 (Foxp3)–PE (all from eBioscience, San Diego, CA, USA). Tissues were stained with Abs against CD4–FITC, p-STAT3–Y705–PE, p-STAT3–S727–PE, p-STAT5-PE, and PIAS3-PE (eBioscience) to examine the population of STAT-expressing cells. The stained tissue sections were visualized on a confocal microscope (LSM 510 Meta; Carl Zeiss, Oberkochen, Germany). Double or triple positive cells were counted in three high-power field (magnification: 200×) per section.