Fig 2: Expression of ESR1 in the trabecular meshwork (TM)/Sclemm’s canal (SC) region of human outflow tissue. ESR1 protein was expressed in the TM/SC region of human outflow tissue (n = 4, 24–60 years of age) taken from both male (a–c) and female (d–f) post-mortem donors. Representative images of H&E (a,d), negative control (b,e), and positive staining (c,f). ESR1, estrogen receptor 1; TM, trabecular meshwork; SC, Schlemm’s canal. Scale bar: 50 µm.

Fig 3: Prioritization of intraocular pressure (IOP)-associated genes differentially expressed in primary open-angle glaucoma (POAG)-affected trabecular meshwork (TM) tissue and/or Schlemm’s canal (SC) cells. Twenty-four IOP-associated genes were differentially expressed (¦FC¦ = 1.5) in POAG-affected TM tissue and/or POAG-affected SC cells (grey circle). Fifteen of the 24 genes were highly expressed (FPKM = 10) in non-glaucomatous TM or SC cells (dashed circle), all of which were located in regions of histone modification. Eight of the IOP-associated variants in/near these 15 genes were located in enhancer or promoter regions (blue circle), and 4 of the 15 were significantly/highly expressed in TM tissue (purple circle). Eight of the 24 genes were specifically expressed in =1 cell cluster defined by single-cell RNA-sequencing of human outflow tissue (red circle), and 2 of the 24 genes were differentially expressed by cyclically stretched human TM cells (green circle). Five of the 24 genes were part of differential correlation pairs (asterisk) and 2 were regulated by ESR1 (two-barred cross).

Fig 4: Effects of APAP and GEN on the expression of estrogen receptors in TM4 cells and PND8 rat Sertoli cells. (A) Gper and Esr1 gene expression in TM4 cells (B) and Esr1 in rat Sertoli cells after 24 h exposure to APAP and GEN alone or mixed at 10, 50, and 100 µM. Data are the means ± SEM of three experiments, each performed in triplicate for each condition. A one-way ANOVA, multiple comparisons; * p = 0.05; ** p = 0.01; *** p = 0.001. Dashed lines show the level of vehicle controls (value of 1.0). (C) Representative pictures of immunofluorescent staining of ER-a (green) in TM4 cells treated with 50 µM of APAP, GEN, or the mixture. DAPI (blue) nuclei staining. Scale: 50 µm.