AMPK Methods and Protocols

Table 1Data obtained from an example experimentCondition dpm

14

C Minus background Palmitate uptake Normalized dpm

3 H Minus background Deoxyglucose uptake Normalized

Zero time

76

38

Control

1590

1514

10.10

372

334

0.81

Control

1625

1549

10.33

1.00

411

373

0.91

1.00

Control

1669

1593

10.63

414

376

0.91

Oligomycin 3065

2989

19.94

735

697

1.69

Oligomycin 3093

3017

20.13

1.94

724

686

1.67

1.97

Oligomycin 3121

3045

20.31

790

752

1.83

AICAR

2401

2325

15.51

420

382

0.93

AICAR

2312

2236

14.92

1.47

406

368

0.89

1.03

AICAR

2381

2305

15.38

399

361

0.88

The described method for substrate uptake measurement was applied. The following values were experimentally determined. Radioactive counts of 20

μL Day Label were

5354 dpm (

14

C) and 14,705 dpm (

3 H). The amount of cells per incubation was 7.0 mg wet mass/mL (~70,000 cardiomyocytes/mL). The duration of the uptake assay was

5 min. Values for palmitate uptake and deoxyglucose uptake are calculated as nmol/(g wet mass

min)

352 Joost J. F. P. Luiken et al.