FIG 1.

Treatment with PF-06409577 results in the phosphorylation of AMPK and ACC. (A) Chemical structure of compound PF-06409577. MW, molecular weight. (B) Immunofluorescence detection of the phosphorylated (Thr 172) catalytic (α) subunit of AMPK (pAMPK) and total AMPK (tAMPK) in Vero cells treated or not treated (control) with PF-06409577 (50 μg/ml) for 24 h. pAMPK or tAMPK are shown in green and cell nuclei in blue. (C) Quantification of the fluorescence of pAMPK and tAMPK in cells treated as described in panel B. (D) Immunofluorescence detection of phosphorylated (Ser79) ACC (pACC) and total ACC (tACC) in cells treated or not treated as described in panel B. pACC and tACC are shown in green and cell nuclei in blue. (E) Quantification of the fluorescence of pACC and tACC in cells treated as described in panel D. (C and E) Data are presented as mean ± SD (n = 20 to 41). Points represent individual cells analyzed. AU, arbitrary units. Bar, 10 μm. In the case of different variances, two-tailed Student's t test P values between control and treated cells were calculated applying Welch's correction. ***, P < 0.005.