Fig 2: Effects of PAR2 inhibitor and ADAM17 inhibitor on the protein levels of acetylated HMGB1, ADAM17 and HB-EGF in cultured mouse epithelial cells. (A) Western blot analysis on the protein levels of acetylated HMGB1, ADAM17 and HB-EGF after mouse epithelial cells were pretreated with FSLLRY-NH2 (100 μM, PAR2 inhibitor) for 24 h and further treated with trypsin for 15 min in the absence or presence of FSLLRY-NH2. (B) The relative HB-EGF level in Figure 4A . (C) The morphology of mouse uteri on day 5 of pregnancy after day 4 pregnant mice were intraluminally injected with 3 μl of saline, 100 μM of FSLLRY-NH2 in saline or 1 μM TAPI-1 in saline at 09:00. (D) The number of implantation sites (blue bands) on day 5 of pregnancy after 3 μl of saline, 100 μM FSLLRY-NH2 in saline, or 1 μM TAPI-1 in saline were intraluminally injected into pregnant mice at 09:00. By injecting 0.1 ml of 1% Chicago sky blue through the tail vein, the implantation sites were counted. (E) The relative ADAM17 in Figure 4A . (F) Western blot analysis on the protein levels of ADAM17 and HB-EGF after mouse epithelial cells were pretreated with TAPI-1 (1 μM, ADAM17 inhibitor) for 30 min and further treated with trypsin for 15 min in the absence or presence of TAPI-1. (G, H) Relative quantitation on the protein levels of HB-EGF and ADAM17 in Figure 4F . *p < 0.05, **p < 0.01, ***p < 0.001.

Fig 3: Effects of lactate, TNFα and trypsin on the levels of HMGB1, acetylated HMGB1, ADAM17 and HB-EGF in cultured mouse epithelial cells. (A) Western blot analysis of the protein levels of HMGB1 and acetylated HMGB1 after epithelial cells were treated with lactic acid for 15 min. (B) Western blot analysis of the protein levels of HMGB1 and acetylated HMGB1 after epithelial cells were treated with TNFα for 15min. (C) Western blot analysis of the protein levels of ADAM17, HMGB1, acetylated HMGB1 and HB-EGF after epithelial cells were treated with trypsin for 15 min. (D-F) Relative quantitative analysis on the protein levels of ADAM17, acetylated HMGB1 and HB-EGF in Figure 2E , respectively. (G) The trypsin level in blastocyst cavity from 100 mouse blastocysts. (H) The trypsin level in the supernatant after 100 mouse blastocysts were cultured in 200 μl of M2 medium for 6 h. **p < 0.01, ***p < 0.001.

Fig 4: Effects of exogenous HMGB1on human decidualization and human uterine epithelial cells. (A) Effects of exogenous HMGB1 on IGFBP1 mRNA level under in vitro decidualization. (B) Western blot analysis on the protein levels of ADAM17, HMGB1, Ace-HMGB1 and HB-EGF after human Ishikawa epithelial cells were treated with trypsin. (C-E) The relative protein levels of ADAM17, acetylated HMGB1 and HB-EGF in Figure 6B , respectively. *p < 0.05; **p < 0.01, ***p < 0.001.

Fig 5: Effects of PAR2 inhibitor and ADAM17 inhibitor on the protein levels of acetylated HMGB1, ADAM17 and HB-EGF after human Ishikawa cells were treated with 500 ng/ml trypsin. (A) Western blot analysis on the protein levels of acetylated HMGB1, ADAM17 and HB-EGF when Ishikawa cells were pretreated with 100 μM FSLLRY-NH2 for 24 h and further treated with 500 ng/ml trypsin in the absence or presence of FSLLRY-NH2. (B, C) The relative protein levels of ADAM17 and HB-EGF in Figure 7A . (D) Western blot analysis on the protein levels of ADAM17 and HB-EGF when Ishikawa cells were pretreated with 1 μM TAPI-1 for 30 min and further treated with 500 ng/ml trypsin for 15 min. (E, F) The relative protein levels of ADAM17 and HB-EGF in Figure 7D .*, p <0.05.