Journal: Advanced Science
Article Title: N‐ glycosylation Modification of CTSD Affects Liver Metastases in Colorectal Cancer
doi: 10.1002/advs.202411740
Figure Lengend Snippet: Identification of N ‐glycosyltransferase for CTSD. A) CTSD with a FLAG label was enriched using FLAG magnetic beads and silver‐stained gel showed that proteins, such as DDOST, were enriched by CTSD. B) 191 proteins were enriched by CTSD N263Q and 235 proteins were enriched by CTSD WT, while 45 proteins exclusively interacted with CTSD WT. C) Expression levels of STT3A, STT3B, and DDOST were upregulated in colon cancer relative to normal tissues. Boxplots show median (central line), upper and lower quartiles (box limits), and 1.5 × interquartile range (whiskers). D) N‐ glycosylated modification at residue 263 of CTSD was regulated by STT3B and DDOST. When STT3B and DDOST were knocked out, molecular weights of CTSD decreased significantly. E) Co‐immunoprecipitation and western blot analysis demonstrated that CTSD formed complexes with STT3B and DDOST. F) Immunofluorescence analysis illustrated the co‐localization of DDOST and STT3B with CTSD, whereas no such co‐localization was observed with STT3A.
Article Snippet: [ ] Anti‐rabbit CTSD (AB‐75811, Abcam), Anti‐mouse CTSD (AB‐302649, Abcam), Anti‐rabbit CTSD (AB‐75852, Abcam), Anti‐mouse GAPDH (60004‐1‐Ig, Proteintech), Anti‐rabbit IgG, HRP‐linked Antibody (Cell Signaling Technology), Anti‐mouse IgG, HRP‐linked Antibody (Cell Signaling Technology), Anti‐mouse ACADM (67742‐1‐Ig, Proteintech), Anti‐rabbit DDOST (14916‐1‐AP, Proteintech), Anti‐mouse STT3A (66581‐1‐Ig, Proteintech), Anti‐mouse STT3B (15323‐1‐AP, Proteintech), Anti‐rabbit ACSL4 (AB‐155282, Abcam), Anti‐rabbit SLC7A11 (AB‐307601, Abcam) and Anti‐rabbit GPX4 (AB‐125066, Abcam) were used.
Techniques: Magnetic Beads, Staining, Expressing, Modification, Residue, Immunoprecipitation, Western Blot, Immunofluorescence
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