Fig 1: Effects of IFA on GALNTs-related gene expression and enzyme activity. (A) Western blot was used to detect the effect of alcohol and IFA pretreatment on the expression levels of GALNTs in the gastric mucosal tissues of rats. (B,C) Determination of GALNT glycosyltransferase activity in GES-1 cells. The color of the control group in black and the alcohol-treated group in red (B). The enzyme activity of GALNTs under different conditions was measured using receptor peptides MUC5AC and MUC6. Black represents the control group, red represents the IFA-only treatment group, blue represents the alcohol treatment group, purple represents the alcohol-treated and low-dose IFA treatment group, and green represents the alcohol and high-dose IFA treatment group (C). * p < 0.05, ** p < 0.01. (D) Molecular docking results of IFA interaction with GALNT1, GALNT2, GALNT10, GALNT12, GALNT14, and GALNT15. (E) Pull-down experiment was utilized to detect the interaction between IFA and GALNT2. rGALNT2 (upper image) and GES-1 cell lysate (lower image) were incubated with IFA-Fe3O4 coupled magnetic beads overnight at 4 °C, and the interaction was detected by GALNT2 antibody.
Fig 2: Effects of IFA on mucin expression in gastric mucosa. (A,B) Statistical map of hexosamine and adhesive mucus content in gastric mucosa. Red represents the control group, blue represents the IFA-only treatment group, green represents the alcohol treatment group, purple represents the alcohol-treated and low-dose IFA treatment group, and black represents the alcohol and high-dose IFA treatment group. *** p < 0.001. (C,D) Real-time PCR was used to detect MUC5A and MUC6 mRNA levels in different gastric mucosa tissues. Black represents the control group, red represents the IFA-only treatment group, green represents the alcohol-treated group, dark blue represents the alcohol-treated and low-dose IFA-treated group, and light blue represents the alcohol-treated and high-dose IFA-treated group. (E) Co-IP assay was used to detect the change in mucin Tn antigen expression. (F) Western blot assay was performed to detect MUC5AC and MUC6 expression levels after the sugar chain of mucin was removed by periodate oxidation-β elimination during the process of extracting gastric mucosal tissue proteins.
Supplier Page from Abcam for Recombinant Human Mucin 5AC protein (GST tag N-Terminus)