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. 2003 Nov 3;198(9):1289–1300. doi: 10.1084/jem.20030057

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Copyright © 2003, The Rockefeller University Press

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Figure 5. — ³⁵Sulfate incorporation and MECA79 reactivity of GlyCAM-1 and CD34. (A and D) Lymph nodes from HEC-GlcNAc6ST−/− and +/+ mice were incubated with Na³⁵SO₄. Immunoprecipitated GlyCAM-1 and CD34 were separated by SDS-PAGE and transferred to PVDF for Western blotting. (A) GlyCAM-1 was detected with an antipeptide Ab (CAMO5). (D) CD34 was detected with an affinity-purified polyclonal Ab. (B and E) ³⁵S-labeled immunoprecipitated ligands GlyCAM-1 (B) and CD34 (E) detected by autoradiography. (C and F) MECA79 reactivity of GlyCAM-1 (C) and CD34 (F) detected by Western blotting. (G) Relative sulfate incorporation and MECA79 reactivity of GlyCAM-1 and CD34 from HEC-GlcNAc6ST−/− mice. ³⁵S incorporation was measured by performing densitometry on autoradiographs as shown in B and E and normalized for protein based on Western blots performed in parallel with protein-specific Abs as in A and D. MECA79 reactivity was measured by performing densitometry on Western blots shown in C and F and normalized based on protein-specific Western blots performed in parallel. Data are the means ± SEMs of three independent preparations.