Fig. 2: Alterations of ADAMTS4 and LRP1 abundance in the human amnion in deliveries with spontaneous rupture of membranes.

a Heatmap of transcriptomic sequencing data displaying alterations in mRNA abundance of ADAMTS family members (ADAMTS1, 4, 5, 9, 15, and 20) possessing proteoglycanase activity toward VCAN in the human amnion in TL (n = 3) as compared to TNL (n = 3). Of these proteoglycanases, only ADAMTS4 manifest significantly increased expression in TL. Blue to red represents expression levels from low to high. b qRT-PCR analysis showing increased ADAMTS4 mRNA abundance in the human amnion in TL (n = 12) compared to TNL (n = 11). c Western blotting analysis showing increased ADAMTS4 and decreased LRP1 protein abundance in the human amnion in TL (n = 7) compared to TNL (n = 7). d qRT-PCR analysis showing decreased LRP1 mRNA abundance in the human amnion in TL (n = 12) as compared to TNL (n = 11). e Single-cell sequencing analysis displaying the cell types expressing ADAMTS4 and LRP1 mRNA in the human amnion. FB, fibroblasts; EpC, epithelial cells; ImC, immunocytes; EpC_FB, epithelial_fibroblasts; ImC_EpC, immune_epithelial cells; ImC_FB, immune_fibroblasts. f Comparison of ADAMTS4 or LRP1 mRNA abundance in cultured human amnion epithelial cells and fibroblasts prepared from TNL (n = 4) as measured with qRT-PCR. AE, epithelial cells; AF, amnion fibroblasts. g Representative immunohistochemical images showing the distribution of ADAMTS4 and LRP1 in the human amnion collected from TNL (n = 3). ADAMTS4 was found mainly in amnion fibroblasts, while LRP1 was found in both amnion fibroblasts and epithelial cells. Scale bars, 25 or 100 μm. Data are mean ± SEM. Statistical analysis was performed with unpaired Student’s t test (b), Mann–Whitney U test (c. d) or paired Student’s t test (f). *p < 0.05, **p < 0.01, ***p < 0.001 vs TNL or AE.