FIGURE 6.

Sgms1 ablation does not impact insulin signaling. C2C12 myotubes exposed to control or the CRISPR/Cas9 knock-out plasmids (SMS1KO) were treated with BSA (control) or BSA-conjugated palmitate (500 μm). A, transcripts encoding Sgms1 and Sgms2 were quantified by qPCR (n = 4). B, data represent the effect of Sgms1 KO on total SMS activity as determined by quantifying the amount of NBD-C₆-ceramide converted from NBD-SM (n = 3). * denotes significance of Sgms1 KO with p value <0.05. C and D, cell lysates were resolved by SDS-PAGE and Western blotted with antibodies recognizing phosphorylated Akt (Ser-473), total Akt, or β-actin (n = 3). * denotes significance of palmitate treatment with a p value of <0.05.