Long-chain saturated fatty acids such as palmitic acid induce insulin resistance and NF-kappaB activation in skeletal muscle cells. Here we investigated the effects of long-chain fatty acid (FA) saturation and configuration on NF-kappaB activity and insulin sensitivity in cultured skeletal muscle cells. Of all tested unsaturated FAs, only elaidic acid (3-fold), cis9,trans11-CLA (3-fold) and trans10,cis12-CLA (13-fold) increased NF-kappaB transactivation in myotubes. This was not accompanied by decreased insulin sensitivity (measured as insulin-induced glucose uptake and GLUT4 translocation). We therefore conclude that FA-induced NF-kappaB activation is not sufficient for the induction of insulin resistance in skeletal muscle cells.