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. 2021 Jun;191(6):1020–1035. doi: 10.1016/j.ajpath.2021.02.018

Figure 6.

Figure 6

Activation of Wnt signaling inhibits dexamethasone (Dex)–induced extracellular matrix in primary human trabecular meshwork (HTM) cells. HTM75OS (A), HTM71FOS (B), and 2019-017 (C) primary HTM cells were treated with 0.1% ethanol (EtOH; vehicle), dimethyl sulfoxide (DMSO; vehicle), 100 nmol/L Dex, 1 μmol/L 6-bromoindirubin-3′-oxime (BIO), 10 μmol/L SB-216763 (SB), and/or 5 μmol/L CHIR99021 (CHIR) for 72 hours. Fibronectin (FN) and collagen-I from conditioned medium (CM) and whole cell lysate (WCL) were used for Western immunoblotting. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as a loading control for WCL. The 2020-001 (D), HTM71FOS (E), and HTM2180 (F) primary HTM cells were treated with 0.1% EtOH (vehicle), DMSO (vehicle), 100 nmol/L Dex, and Dex + glycogen synthase kinase 3β inhibitors (1 μmol/L BIO, 10 μmol/L SB, and/or 5 μmol/L CHIR) for 12 to 14 days, and were immunostained with anti–EDA-FN or EDB-FN antibodies (green). Nuclei were stained using Hoechst-33342 and/or DAPI (blue). Scale bar = 20 μm (D–F).