- + - +

pIRS1(S636/639)

IRS1

actin

PF4708671

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.

Supplemental Figure S2

HIF1α

actin

- + - + - + - + -

o/n chronic

Rap

IR

insulin

DMEM MEM

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 beforestimulation.