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. 2008 Apr 22;118(5):1815–1824. doi: 10.1172/JCI32467

Figure 10. LH from GalNAc-4-ST1–/– mice has increased levels of Siaα2,6GalNAc.

Figure 10

Extracts were prepared from 5 WT and 5 GalNAc-4-ST1–/– mice that had not been castrated (A and B) or had been castrated (cast) (C–F). Identical aliquots of each extract were incubated with immobilized WFA or SNA-1, which binds structures terminating with GalNAc or Siaα2,6GalNAc, respectively. Terminal sialic acid was removed by digestion with neuraminidase as indicated prior to incubation with WFA or SNA-1 (B, D, and F). Equal aliquots of the starting material (In), the unbound fraction (Ub), and the fractions selectively eluted with GalNAc or lactose (Elution) were analyzed by SDS-PAGE and Western blotting with anti-LHβ. (A) Binding of pituitary extracts by WFA-agarose. (B) Binding of pituitary extracts by WFA-agarose following neuraminidase digestion. (C) Binding of pituitary extracts from castrated mice by WFA-agarose. (D) Binding of pituitary extracts from castrated mice by WFA-agarose following neuraminidase digestion. (E) Binding by SNA-1–agarose. (F) Binding by SNA-1–agarose following digestion with neuraminidase.