Research Paper Volume 13, Issue 17 pp 21547—21570

HSP90 acts as a senomorphic target in senescent retinal pigmental epithelial cells

Heat shock protein 90 inhibitor IPI-504 suppresses the expression and secretion of senescence-associated inflammatory factors in senescent ARPE-19. (A–D) quantitative PCR measuring the expression of IL-1β, IL-6, IL-8, MCP-1 in day-4 senescent ARPE cells treated with media containing PBS (0) or IPI-504 at the concentrations of 0.2, 0.5, 1 and 5 μM. (E–F) ELISA assay measuring the secretion of IL-1β and IL-8 in the supernatants of senescent ARPE-19 cells treated with IPI-504. con: proliferative ARPE-19 cells, SC: senescent ARPE-19 cells (day-4), SC+IPI504: Day-4 SC cells were treated with 1 μM IPI-504 for 24 hours. (G) Immunoblot the expression of IL-1β, cleaved caspase 1, NLRP3 and GAPDH in control ARPE-19 cells (lane 1), or day-4 senescent ARPE-19 cells treated with IPI-504 at 0, 1 and 5 μM (lanes 2, 3 and 4). The data in each quantitation figure were collected from four independent experiments (n = 4). The two-tailed unpaired t-test was used for statistical analysis, *p ***p

Figure 3. Heat shock protein 90 inhibitor IPI-504 suppresses the expression and secretion of senescence-associated inflammatory factors in senescent ARPE-19. (AD) quantitative PCR measuring the expression of IL-1β, IL-6, IL-8, MCP-1 in day-4 senescent ARPE cells treated with media containing PBS (0) or IPI-504 at the concentrations of 0.2, 0.5, 1 and 5 μM. (EF) ELISA assay measuring the secretion of IL-1β and IL-8 in the supernatants of senescent ARPE-19 cells treated with IPI-504. con: proliferative ARPE-19 cells, SC: senescent ARPE-19 cells (day-4), SC+IPI504: Day-4 SC cells were treated with 1 μM IPI-504 for 24 hours. (G) Immunoblot the expression of IL-1β, cleaved caspase 1, NLRP3 and GAPDH in control ARPE-19 cells (lane 1), or day-4 senescent ARPE-19 cells treated with IPI-504 at 0, 1 and 5 μM (lanes 2, 3 and 4). The data in each quantitation figure were collected from four independent experiments (n = 4). The two-tailed unpaired t-test was used for statistical analysis, *p < 0.05, ***p < 0.001.