Journal: The Journal of Biological Chemistry

Article Title: Enzymatic basis of branching and extension of O -Man glycans for keratan sulfate biosynthesis

doi: 10.1016/j.jbc.2026.111140

Figure Lengend Snippet: Enzyme assays of KS biosynthetic enzymes toward linear and branched O -Man glycans . A , enzymes involved in the KS biosynthesis. B , qPCR analysis of B4galt1 , B4galt4 , B3gnt7 , Chst1 , Chst2 and Chst5 mRNAs in GnT-IX-KO mouse brains (P0) was performed, and their relative levels normalized by that of Gapdh are shown as a graph (n = 3, mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, unpaired t test). C , soluble B4GALT1, B4GALT4, B3GNT7, CHST1, CHST2, and CHST6 were expressed in HEK293 T cells and purified from the media using a Ni2+ column. The purified enzymes were separated by SDS-PAGE and visualized by CBB staining. D – F , Purified B4GALT1, B4GALT4, B3GNT7, CHST1, CHST2, and CHST6 were incubated with GnM-S-Flu or Gn(Gn)M-S-Flu (for B4GALT1 and B4GALT4) ( D ), GalGnM-S-Flu or GalGn(GalGn)M-S-Flu (for B3GNT7) ( E ), or GnGalGnM-S-Flu or GnGalGn(GnGalGn)M-S-Flu (for CHSTs) ( F ), and the reaction mixtures were analyzed by reverse-phase HPLC. The amount of CHST1 used for GnGalGn(GnGalGn)M-S-Flu was one-20th of the amount for GnGalGnM-S-Flu, while the same amounts of the other enzymes were used for linear and branched substrates. The positions of sulfate residue were predicted by the substrate specificity of the enzymes. The specific activities were calculated from the peak areas (n = 8–17, mean ± SD, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001, Holm-Sidak’s multiple comparison test for B4GALT1, B4GALT4, CHST1, CHST2, and CHST6, and unpaired t test for B3GNT7).

Article Snippet: The primers and probes were purchased from Applied Biosystems as follows: Mm00480752_m1 for B4galt1 Mm00480087_m1 for B4galt4 , Mm01295423_m1 for B3gnt7 , Mm01334076_m1 for Chst1 , Mm00490018_g1 for Chst2 , Mm00517342_m1 for Chst5 , and 4,308,313 (TaqMan Rodent GAPDH Control Reagents) for Gapdh .

Techniques: Purification, SDS Page, Staining, Incubation, Residue, Comparison

Journal: The Journal of Biological Chemistry

Article Title: Enzymatic basis of branching and extension of O -Man glycans for keratan sulfate biosynthesis

doi: 10.1016/j.jbc.2026.111140

Figure Lengend Snippet: Actions of KS biosynthetic enzymes on O -Man glycans . B4GALT1, B4GALT4, and CHST1 preferentially act on GnT-IX-produced branched O -Man glycans.

Article Snippet: The primers and probes were purchased from Applied Biosystems as follows: Mm00480752_m1 for B4galt1 Mm00480087_m1 for B4galt4 , Mm01295423_m1 for B3gnt7 , Mm01334076_m1 for Chst1 , Mm00490018_g1 for Chst2 , Mm00517342_m1 for Chst5 , and 4,308,313 (TaqMan Rodent GAPDH Control Reagents) for Gapdh .

Techniques: Produced

Journal: International Journal of Molecular Sciences

Article Title: Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide Particles

doi: 10.3390/ijms241814253

Figure Lengend Snippet: Function of the genes examined in the study.

Article Snippet: B4GALT1 , UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase, polypeptide 1 , Rn01764643_m1 , Hs00155245_m1.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Uremic Toxin-Induced Exosome-like Extracellular Vesicles Contain Enhanced Levels of Sulfated Glycosaminoglycans which Facilitate the Interaction with Very Small Superparamagnetic Iron Oxide Particles

doi: 10.3390/ijms241814253

Figure Lengend Snippet: List of applied TaqMan™ probes (Thermo Fisher) in the gene expression experiments.

Article Snippet: B4GALT1 , UDP-Gal:betaGlcNAc beta 1,4-galactosyltransferase, polypeptide 1 , Rn01764643_m1 , Hs00155245_m1.

Techniques: Gene Expression