Inactivation of α1AMPK increases basal and stearate-stimulated TNF-α mRNA. A and B, establishment of a macrophage cell line with α1AMPK knockdown (α1KD). RAW264.7 macrophages were infected with the α1AMPK shRNA lentivirus or control lentivirus, and selected with puromycin for 8 days. α1AMPK RNA (A) and protein (B) levels were evaluated by real time RT-PCR and immunoblotting, respectively. C and D, α1AMPK knockdown increases basal and stearate- or LPS-stimulated TNF-α mRNA. The α1AMPK-knockdown and control macrophages were treated with vehicle (BSA for stearate and H2O for LPS), stearate (500 μm), or LPS (100 ng/ml) for 4 h. E and F, inactivation of α1AMPK by DN-α1AMPK overexpression increases basal and stearate- or LPS-stimulated TNF-α mRNA. RAW264.7 macrophages were transfected with pcDNA3.1 or DN-α1AMPK, and then treated with vehicle (BSA for stearate and H2O for LPS), LPS (100 ng/ml), or stearate (500 μm) as indicated for 4 h. TNF-α mRNA was measured by real time RT-PCR. Data are expressed as mean ± S.E., n = 6. *, p < 0.05. A.U., arbitrary units.