A

PKC-pTyr311

PKC

0 0.5 1 2 Thr (min)

B

C

D

DMSO

- +

Rott LY

eNOS-pSer1179

eNOS

- + - + Thr

- +

Basal A23

eNOS-pSer1179

eNOS

Rott - +

eNO

S-p

(%

)

- +

- - pre co

Thr

eNOS-pSer1179

eNOS

eNOS-pThr497

PMA

+ +

eNO

S-p

(%

) †

On Line Figure I. A. HUVECs pretreated with 10 mol/L rottlerin (Rott), 10 mol/L LY294002 (LY), or control vehicle (0.1% DMSO) for 30 min were stimulated with 10 U/mL thrombin (Thr) for 1 min. B. BAECs were stimulated with thrombin for the indicated time periods. PKC Tyr311 phosphorylation was evaluated by immunoblotting. C. BAECs pretreated with or without 10 mol/L rottlerin (Rott) for 30 min were stimulated with 10 mol/L A23187 for 1 min. D. BAECs co-treated (co) or pretreated (pre 5 min) with 1 mol/L PMA were stimulated with 10 U/mL thrombin for 1 min. These data shown are representative of 3 separate experiments giving similar results (mean + SEM. *p < 0.05 compared to basal. †p < 0.05 compared to stimulated control.).

†

0

25

50

75

100

pSer1179

pThr497†

0 25

50 75

100 125 150

eNO

S-p

(%

)

†

**

0

25

50

75

100

125