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This study investigates the impact of PF4708671 and rapamycin on insulin signaling in retinal pigment epithelial (RPE) cells. Cells were incubated in serum-free DMEM with high (25 mM) or low (5.5 mM) glucose, treated with 10 µM PF4708671, and stimulated with 1 µg/ml insulin. Immunoblot analyses revealed changes in insulin receptor signaling and HIF1α expression. Our findings suggest significant interactions between insulin signaling pathways and pharmacological agents, providing insight into potential therapeutic avenues for metabolic regulation in retinal cells.
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PF4708671 - + - + pIRS1(S636/639) IRS1 actin Supplemental Figure S1 Immunoblot analysis. RPE cells were incubated in serum free DMEM (25 mM glucose) in the presence or absence of 10 µM PF 4708671 for 42-44 hours and then were stimulated with 1 µg/ml insulin and lysed.
Rap IR o/n chronic - + - + - + - + - insulin HIF1α actin DMEM MEM Supplemental Figure S2 Immunoblot analysis. RPE cells were maintained either in DMEM (25 mM glucose) or in MEM (5.5 mM glucose). Cells were incubated in serum-free respective medium for 42-44 hr and then stimulated with 1 µg/ml insulin for 15 min and lysed, Rap – 10 nM rapamycin; o/n – cells were pretreated with rapamycin overnight before stimulation; chronic – cells pre-treated with rapamycin for 2 weeks before stimulation; IR – cells were pre-treated with 1 µg/ml insulin overnight before stimulation.
