Fig 2: Matrigel and Transwell assays to detect the effect of PAR-2 shRNA on EC109 cell invasion and migration. Cells were stained using hematoxylin and eosin. (A) Cell invasion analysis was measured by Matrigel method on the EC109 cell and PAR-2 shRNA groups (magnification, ×200). (B) Cell migration analysis was measured by Transwell method on EC109 cells group and PAR-2 shRNA group (magnification, ×200). PAR-2, protease-activated receptor 2; shRNA, small hairpin RNA.

Fig 3: Tube formation ability of EPCs. Tryptase promoted the tube formation ability of EPCs; a PAR-2 agonist had a similar effect to tryptase, while tryptase inhibitor APC366 or PAR-2 inhibitor SAM 11 inhibited the effect of tryptase on tube formation Magnification, ×100. EPCs, endothelial progenitor cells; PAR-2, proteinase activated receptor-2.

Fig 4: Effects of EEDAR on signaling molecules that regulate TJ structure and function in spleen-deficiency diarrhea rats (n = 3). The phosphorylation levels of p38 and PAR-2 in total colon homogenates were measured by Western blot analysis (A,B). Representative images of immunohistochemistry sections (200× magnification) from PAR-2 and P-P38 (C) colons indicated different pathological changes in spleen deficiency diarrhea rats treated with EEDAR-L (1.85 g·kg-1), EEDAR-H (7.40 g·kg-1), and SLBZS. Total protein levels of P-MLC and MLCK were measured by Western blot analysis (E,F). Bands were quantified and values normalized to non-phosphorylated protein or ß-actin levels. Values represent means ± SEM. # p < 0.05 vs. Control; * p < 0.05 vs. Model. (Scale bar 1:100).

Fig 5: DPM and PAE modulated PAR-2 expression in opposite ways. (a) Western blot showed a robust increase in the PAR-2 protein caused by DPM in HaCaT cells, and the treatment with PAE was able to reverse the effect of DPM in a concentration-dependent manner. (b) Transcriptional regulation of PAR-2 was demonstrated to be upregulated by DPM, which could be inhibited by PAE. (* p < 0.05, *** p < 0.001).