AMPK Methods and Protocols

2.2 Reagents
and Materials

1. Dimethyl sulfoxide (DMSO).


2. Oligomycin stock solution: 100μM oligomycin in DMSO.
   Prepare a 10 mM oligomycin solution by dissolving 5 mg of
   oligomycin in 633μL of DMSO. Make several aliquots to avoid
   freeze-thaw cycles. Further dilute the 10 mM oligomycin

0

50

100

0

50

100

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50

100

CD4

CD8

LB

WT

AMPK KO-T

% viabl

ecells

Oligo

Oligo

Oligo

% viable cells

% viable cells

Fig. 3AMPK-deficient T lymphocytes are more susceptible to metabolic

mitochondrial stress. Splenocytes (10^6 cells in 500 μL of culture medium)

were incubated for 6 h with graded doses of oligomycin (1.1, 3.3, and 10 nM),

with the DMSO solvent or with medium. The relative numbers of viable CD4+,

CD8+, and B cells in each experimental sample were estimated as described in

Fig. 2. Results are expressed as % of viable cells found in the control (medium

alone) group

Flow Cytometry Analysis of Cell Viability upon Metabolic Stress 469