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Figure 3. Establishing an SI-induced oxidative stress model. (A) Baseline mRNA expression of complement genes CFH and C3 as well as NRF2 response genes HMOX1 and NQO1 were analyzed by quantitative reverse transcriptase polymerase chain reaction and data from four independent batches of cells (with three technical replicates per batch) were pooled. Data were normalized against HPRT1 expression and calibrated against the mean of all LR cell lines (n = 4). (B) iPSC-RPE cultured in 96-well plates for four weeks were subjected to 24 h treatment with 0.5 or 3 mM SI. Relative cytotoxicity in relation to untreated cells was determined using an LDH release assay and was not increased upon treatment with 0.5 mM SI while increased upon treatment with 3 mM SI, which served as a positive control. Data are presented as mean + standard deviation (SD) (n = 4). (C) ZO-1 stainings visualize iPSC-RPE monolayer integrity after treatment with 0.5 mM SI for 24 h and show no impact of the treatment on monolayer integrity and cell morphology (scale bar: 20 µm). (D) TEER measurements were performed to confirm monolayer integrity and showed no 24 h SI treatment-dependent changes on HR or LR cell lines (n = 4). Statistical significance was tested using a Mann-Whitney U-test. SI: Sodium iodate; iPSC: induced pluripotent stem cell; TEER: transepithelial electrical resistance; RPE: retinal pigment epithelium; HR: high-risk; LR: low-risk.