Table 1.
Primers used for quantitative real-time PCR.
Fig 1.
Effect of omidenepag (OMD) on human trabecular meshwork (HTM) cell viability.
HTM cells were treated with vehicle or OMD (10 nM, 100 nM, and 1 μM) for 24 h, and cell viability was assessed using MTS assays. (a) Typical phase-contrast microscopic appearance of HTM cells after treatment. Scale bars = 200 μm. (b) Absorbance in each well was normalized to that in the vehicle-treated wells and presented as percentages. Each value represents the mean ± S.E. (n = 6). *P < 0.05, compared with the vehicle-treated group by Dunnett’s multiple comparison test. MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium.
Fig 2.
Effect of omidenepag (OMD) on extracellular matrix (ECM) gene expression in 2D- and 3D-cultured human trabecular meshwork (HTM) cells.
2D- or 3D-cultured HTM cells were treated with vehicle, 10 nM, 100 nM, or 1 μM OMD for 6 or 24 h, and the gene expression changes in ECMs [(a) FN1, COL1A1, COL1A2, and COL4A2, (b) COL12A1, COL13A1, and COL18A1] were assessed using quantitative real-time PCR. The mRNA expression level in each gene was normalized to that for GAPDH and presented as percentages relative to the vehicle-treated group. Each value represents the mean ± S.E. (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 compared with the vehicle-treated group by Dunnett’s multiple comparison test.
Table 2.
P values for the gene expression changes in OMD-treated groups compared with the vehicle-treated group.
Fig 3.
Effect of omidenepag (OMD) on matrix metalloproteinase (MMP) gene expression in 2D- and 3D-cultured human trabecular meshwork (HTM) cells.
2D- or 3D-cultured HTM cells were treated with vehicle, 10 nM, 100 nM, or 1 μM OMD for 6 or 24 h, and the gene expression changes of MMPs [(a) MMP1, MMP2, MMP3, and MMP9, (b) MMP11, MMP12, MMP14, and MMP15, (c) MMP16, MMP17, and MMP24] were assessed using quantitative real-time PCR; gene expression was assessed using real-time PCR. The mRNA expression level in each gene was normalized to that for GAPDH and presented as percentages relative to the vehicle-treated group. Each value represents the mean ± S.E. (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 compared with the vehicle-treated group by Dunnett’s multiple comparison test.
Fig 4.
Effect of omidenepag (OMD) on tissue inhibitors of metalloproteinase (TIMP) gene expression in 2D- and 3D-cultured human trabecular meshwork (HTM) cells.
2D- or 3D-cultured HTM cells were treated with vehicle, 10 nM, 100 nM, or 1 μM OMD for 6 or 24 h, and the gene expression changes of TIMPs (TIMP1, TIMP2, TIMP3, and TIMP4) were assessed using quantitative real-time PCR. The mRNA expression level in each gene was normalized to that of GAPDH and presented as percentages relative to the vehicle-treated group. Each value represents the mean ± S.E. (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 compared with the vehicle-treated group by Dunnett’s multiple comparison test.