AMPK Methods and Protocols

example, 2-deoxyglucose and ionomycin have been used to glob-

ally activate AMPK through activation of LKB1 and CaMKKβ,

respectively, and Compound C is used for global inhibition of

AMPK. Due to the global nature of these perturbations, these

AMPK

Biosensors

Organelle

markers Merged

Organelle

markers Merged

Mitochondria

Golgi apparatus

Lysosome

Nucleus

ER

Cytosol

Plasma membrane

PM-ABKAR

Nuc-ABKAR

Cyto-ABKAR

ER-ABKAR

Mito-ABKAR

Golgi-ABKAR

Lyso-ABKAR

Diffusive

Cytosol

Nucleus

Plasma membrane

Golgi apparatus

ER

Mitochondria

Lysosome

OTS: LYN

OTS: The SV40 large T-antigen

OTS: CYP2C1

OTS: LAMP1 OTS: AKAP1

OTS: NOS3

OTS: HIV-Rev

AMPK

Biosensors

ABKAR Hoechst33342

Cyto-ABKAR Hoechst33342

Nuc-ABKAR Hoechst33342

PM-ABKAR LYN

Golgi-ABKAR Giantin

ER-ABKAR CB5

Mito-ABKAR Mitotracker red

Lyso-ABKAR Lysotracker red

Fig. 2The localization of organelle-specific ABKARs. Scale bar, 10μm. Reproduced from [11] with permission

260 Takafumi Miyamoto et al.