Orlistat inhibits AMPK activation without affecting cAMP levels and PKA
signaling. A, intracellular levels of cAMP in 3T3-L1 adipocytes
treated with forskolin with/without orlistat as described in
Fig. 2. B, cell
lysates obtained in the experiment described in
Fig. 2 were immunoblotted
(IB) with antibodies recognizing phosphorylated serine and threonine
residues of PKA substrates (∼P-PKA), totalα-AMPK, the PKA
downstream targets P-LKB1 Ser-431, P-CREB Ser-133, and β-actin.
Densitometric analyses are as follows: C, P-PKA substrate ∼62
kDa; D, α-AMPK; E, P-LKB1 Ser-431; and F,
P-CREB Ser-133. Immunoblots shown are representative of nine samples obtained
in three independent experiments. Results in A, C, D, E, and
F are expressed as means ± S.E. (n = 9).
Significantly different from control group: ***, p <
0.001. Iso, isoproterenol.