1.
pIRS1(S636/639)
IRS1
actin PF
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 for hours and then were stimulated with 1 µg/ml insulin and lysed.

2. Rap IR insulin - + - + - + - + - HIF1α actin DMEM MEM
o/n
chronic IR
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 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.