FIGURE 6.

HGA induced the association of IRS-1 with PKCα and RAGE in L6 cells. a, L6 skeletal muscle cells were treated with 0.1 mg/ml HA or HGA for 24 h. Cells were lysed as described under “Experimental Procedures,” and lysates were then precipitated (IP) with selective anti-RAGE antibodies and loaded on SDS-PAGE. Gels were then transferred onto nitrocellulose filters and incubated with recombinant biotinylated PKCα (OB PKCα) as described under “Experimental Procedures” (left panel). Cell lysates precipitated with selective anti-RAGE antibodies were also immunoblotted (WB) with PKCα antibody (right panel). The autoradiographs shown are representative of three independent experiments. b, lysates from L6 skeletal muscle cells treated as above were precipitated with pre-immune serum (PI) or selective anti-RAGE antibodies and loaded on SDS-PAGE. Gels were then transferred on nitrocellulose filters and immunoblotted with anti-IRS1, anti-PKCα, or anti-RAGE antibodies. The autoradiographs shown are representative of three independent experiments.