Fig. 4: Combined supplementation of FAC and CSE to the culture media is sufficient to decrease both uptake and degradation of POS by hiPSC-RPE cells. | Cell Death Discovery

Fig. 4: Combined supplementation of FAC and CSE to the culture media is sufficient to decrease both uptake and degradation of POS by hiPSC-RPE cells.

From: Environmental stress impairs photoreceptor outer segment (POS) phagocytosis and degradation and induces autofluorescent material accumulation in hiPSC-RPE cells

Fig. 4

a–c Representative western blot images (a) and corresponding quantitative analyses (b, c) showing reduced uptake of RHO/POS at the 0 h time point (a, b) and decreased degradation of ingested RHO/POS at 24 h time point (a, c) in untreated versus FAC + CSE-treated (200 µg/ml + 0.5%) hiPSC-RPE cultures. Data are presented as mean ± SEM, n = 3 independent trials. *P ≤ 0.05. ACTN served as loading control. d–e Representative western blot images (d) and corresponding quantitative analyses (e) displaying reduced levels of active CTSD (32 kDa) in POS-fed and FAC + CSE-treated hiPSC-RPE cultures compared to only POS-fed hiPSC-RPE cultures. Data are presented as mean ± SEM, n = 3 independent trials. *P ≤ 0.05, ***P ≤ 0.001. Note: ACTN served as loading control for western blot analyses. f Representative fluorescent microscopy (top panel) and confocal microscopy images (bottom panel) demonstrating equivalent staining of calcein-AM-positive live cells (top panel) and ZO-1 localization (bottom panel) in untreated and FAC + CSE-treated (200 µg/ml + 0.5%, 24 h) hiPSC-RPE cultures (scale bar = 20 µm). g TER ≥ 150 Ω cm2 was seen at both baseline and 24 h time point in untreated and FAC + CSE-treated (200 µg/ml + 0.5%, 24 h) hiPSC-RPE cultures. Data are presented as mean ± SEM. n = 3 independent trials

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