Fig 2: ET-1 upregulates Fibronectin and a-SMA expression through ETAR/ETBR. Representative images of IF analysis of MCs stimulated with ET-1 and/or BQ123+BQ788 for 48 h. (A) Cells are stained for F-actin (green), Fibronectin (red), and DAPI (blue) or (B) for F-Actin (green), a-SMA (red) and DAPI (blue). Bars, 20 µm.

Fig 3: Increased infiltration of F4/80 positive macrophages is associated with both ETAR and ETBR expression in cerulein induced acute inflammation: Dual color immunofluorescence images of the pancreatic sections stained for ETAR and ETBR (red) and macrophage marker F4/80 (green) following cerulein and saline treatment mice at days 0, 2, 7 and 21. The representative images for ETAR (A) and ETBR (B) are depicted in panels A and C, respectively, with areas showing co-localization of ET-receptors with macrophage markers highlighted in box and zoomed in inset. The degree of overlap between F4/80 positive macrophages and ETAR or ETBR was measured using ImageJ using Manders overlap coefficients (B and D respectively). (** p < 0.005, ns = not significant) (Scale bar = 20 µm; zoom scale bar = 10 µm).

Fig 4: EDNRB protein expression in breast cancer cell lines. (A) Representative Western blot showing EDNRB protein expression in HMEC, MCF-7, MDA-MB-231 and BT-549 cell lines; (B) Semi-quantitative analysis of EDNRB relative to beta-actin (ACTB) in 3 biological replicates (2-way ANOVA p<0.0001); (C) representative flow cytometry charts showing EDNRB cell-surface expression in HMECs (top), MCF7 (middle), and MDA-MB-231 (bottom) cell lines.

Fig 5: Analysis of ETRs expression in murine schistosomiasis.(A) Micrographs of livers and spleens from different weeks after infection. Scale bar, 1 cm. (B) H&E, Masson’s trichrome and Sirius Red staining of liver sections. Scale bar, 200 µm. (C) Collagen content in livers determined as hydroxyproline content (n = 6). (D) Granuloma size measured from H&E-stained liver sections(n = 6). (E) Fibrosis scores measured from Masson’s trichrome staining of liver sections (n = 6). (F) Positive staining areas for Sirius Red were measured using IPP software (n = 6). (G) H&E and Masson’s trichrome staining of spleen sections. Insets show a higher magnification of the outlined area. Black arrows indicate the Trabeculae. Scale bar, 200 µm. (H) Representative immunohistochemical staining for ETAR and ETBR in the infected spleens. Insets show a higher magnification of the outlined area. Black arrows indicate the ETRs positive cells. Scale bar, 100 µm. (I-N) The expression of ET-1, ETAR, ETBR, Col1a1, Col3a1 and IL-10 in spleens during infection was detected by qPCR (n = 3–6). (O) The IL-10 concentration in spleen tissue homogenates was determined by ELISA (n = 6). (P-R) ETAR and ETBR proteins were determined by western blotting, quantified using Image J analysis, and normalized to GAPDH (n = 5). Data are represented as mean ± SEM. All data are representative of at least three independent experiments. Significance was determined by the two-tailed Student’s t test (C-F, I-O, Q-R). *P < 0.05, **P< 0.01, ***P< 0.001, ****P< 0.0001, compared with 0W samples (C, F, I-O, Q-R) and compared with 4W samples (D-E).