Fig. 5.

The GS–GN³⁴ interaction in mammalian cells. (A) COS1 cells were transfected with the indicated GST-MmGN1 constructs in pEBG6P or MmGS2 (untagged) constructs in pCMV5. GST-MmGN1 and MmGS2 transfected lysates were mixed, and complexes were captured on glutathione resin and analyzed by immunoblotting with the indicated antibodies. Results are representative of three independent experiments. UTF, untransfected. (B) COS1 cells were transfected with the indicated MmGS2 constructs in pCMV5 vector. MmGS2 was captured with bacterial GST-MmGN1 on glutathione resin and immunoblotted with the indicated antibodies. Input lysates were adjusted for differences in MmGS2 expression. Results are representative of three independent experiments. UTF, untransfected. (C) As in B, except that each GS mutation was combined with a Ser8Ala phosphorylation site mutation.