Fig 1: The effect of TGF-β1 on pro-MMP-2 and pro-MMP-9 gelatinolytic activity in equine endometrial fibroblasts and epithelial cells. The effect of TGF-β1 (5 ng/ml) on pro-MMP-2 and pro-MMP-9 gelatinolytic activity in endometrial epithelial cells (A,B) (n = 5) and fibroblasts (C–F; n = 6) cultured in vitro. The dotted line indicates the place where blots from the same gel were grouped. All values are expressed as a fold change. Asterisks indicate statistical differences (*P < 0.05) from the respective control, as determined by a nonparametric Mann-Whitney U test.
Fig 2: Relative mRNA transcription of matrix metallopeptidase 2 (MMP2) (A,B) and MMP9 (C,D) in follicular phase (FP) and mid-luteal phase (MLP) mare endometrial explants treated for 24 or 48 h with medium alone (control), elastase (ELA: 0.5 μg/mL), sivelestat (SIV: 10 μg/mL), or ELA (0.5 μg/mL) + SIV (10 μg/mL). Data are shown as median with interquartile range. Results were considered significant at p < 0.05. Different superscript letters indicate significant differences between treatments within each treatment time (a,b—24 h; x,y—48 h). Asterisks indicate statistical differences between times of treatment for the same treatment (* p < 0.05).
Fig 3: The effect of TGF-β1 on MMP-1, -2, -3, and -9 in equine endometrial fibroblasts. The effect of TGF-β1 (5 ng/ml) on Mmp1 mRNA transcription (A) and MMP-1 secretion (B), Mmp2 mRNA transcription (C) and MMP-2 secretion (D), Mmp3 mRNA transcription (E) and MMP-3 secretion (F), and Mmp9 mRNA transcription (G) and MMP-9 secretion (H) from endometrial fibroblasts cultured in vitro (n = 6) for 24 h and 48 h. All values are expressed as a fold change. Asterisks indicate statistical differences (*P < 0.05) from the respective control, as determined by a nonparametric Mann-Whitney U test.
Fig 4: The effect of TGF-β1 on MMP-1, -2, -3, and -9 in equine endometrial epithelial cells. The effect of TGF-β1 (5 ng/ml) on Mmp1 mRNA transcription (A) and MMP-1 secretion (B), Mmp2 mRNA transcription (C) and MMP-2 secretion (D), Mmp3 mRNA transcription (E) and MMP-3 secretion (F), and Mmp9 mRNA transcription (G) and MMP-9 secretion (H) from endometrial epithelial cells cultured in vitro (n = 5) for 24 and 48 h. All values are expressed as a fold change. Asterisks indicate statistical differences (*P < 0.05) from the respective control, as determined by a nonparametric Mann-Whitney U test.
Fig 5: Expression of MMP-3 and -9 in endometrium during mare endometrial fibrosis. Endometrial Mmp3 mRNA transcription (A) and MMP-3 concentration (B) and Mmp9 mRNA transcription (C) and MMP-9 concentration (D) in the midluteal phase and follicular phase of the estrous cycle in the progression of mare endometrial fibrosis (Kenney and Doig’s endometrium categories I, IIA, IIB and III) in equine endometrium. Asterisks indicate statistical differences between Mmp3 and Mmp9 mRNA transcription/protein expression during mare endometrial fibrosis, within the midluteal or follicular phases (*P < 0.05; **P < 0.01).
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