Figure 2. EMT is associated with alteration in glycosylation in PRH.

(A) Normal phase HPLC chromatogram of PRH associated N-linked glycan at 24 and 96 hours post-culture. (B) The same samples after treatment with Arthrobacter ureafaciens sialidase to remove sialic acid. The core fucosylated bi-antennary and tri-antennary glycans that are increased at the 96 hour time point have glucose unit (GU) values of 7.55 and 8.30 and are indicated with an arrow in the 96 hour time point. For panels A and B glucose values are provided for the major peaks and the GU ladder is provided along the X-axis. Y-axis for these panels represents the fluorescent intensity of glycan. (C,D) Lectin blot analysis of independently isolated PRH at 24, 48, 72, 96 or 120 hours post culture. 10 μg of total cellular protein was resolved by SDS-PAGE and sialylated protein (C) or fucosylated protein (D) detected by lectin blotting using the Sambucus nigra lectin (SNA), which binds to α-2,6 and to a lesser degree to α-2,3 linked sialic acid or the fucose binding lectin N224Q rAAL lectin (see Methods). In panel C, specific bands are shown to be increased are indicated by the asterisk.