Journal: Future Science OA

Article Title: Glucosamine induces apoptosis of cholangiocarcinoma cells by suppressing high-mannose type N -glycosylation and EGFR/STAT3 signaling

doi: 10.1080/20565623.2026.2641244

Figure Lengend Snippet: Glucosamine (GlcN) interrupted intracellular signaling and suppressed glycosylation. (A) Glucosamine treatment resulted in a reduced molecular weight of epidermal growth factor receptor (EGFR). This led to (B) suppression of phosphorylation of signal transducer and activator of transcription 3 (STAT3) at Y705. (D) The Western blots demonstrated the molecular weight reduction of glycoprotein 130 (gp130), suggesting glucosamine inhibits the glycosylation of intracellular proteins. (D) Concanavalin A (con A) lectin blots confirmed that glucosamine can inhibit the global N -glycosylation of the intracellular proteins in CCA cells. Western blots are representative of three biological replications with the same trend of results. Graphs show the averaged band intensities of proteins from three biological replications. GAPDH was used as a loading control, and the control groups (0 mM glucosamine) were assigned a factor of 1. * p < .05, ** p < .01, *** p < .001.

Article Snippet: Membranes were blocked with 5% skim milk (w/v) in Tris-buffered saline with 0.1% Tween 20 (TBST), and then incubated with primary antibodies included anti-X-linked inhibitor of apoptosis protein (XIAP) (Santa Cruz Biotechnology, Dallas, TX), anti-cyclin D1 (Cell Signaling, Cambridge, MA), anti-p21 (Cell Signaling), anti-caspase 9 (Cell Signaling), anti-caspase 3 (Cell Signaling), anti-cleaved caspase 3 (Cell Signaling), anti-Poly (ADP-ribose) polymerase (PARP) (Cell signaling), anti-cleaved PARP (cell signaling), anti-cyclin dependent kinase (CDK) 4 (Proteintech, Rosemont, IL), anti-CDK6 (Proteintech), anti-epidermal growth factor receptor (EGFR) (Cell Signaling), anti-signal transducer and activator of transcription 3 (STAT3) (Cell Signaling), anti-pSTAT3 (Y075) (Cell Signaling), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (EMD Millipore, Darmstadt, Germany).

Techniques: Glycoproteomics, Molecular Weight, Phospho-proteomics, Western Blot, Control

Journal: Future Science OA

Article Title: Glucosamine induces apoptosis of cholangiocarcinoma cells by suppressing high-mannose type N -glycosylation and EGFR/STAT3 signaling

doi: 10.1080/20565623.2026.2641244

Figure Lengend Snippet: High glucose (Glc) rescued glucosamine (GlcN)-treated cholangiocarcinoma (CCA) cells. (A) High glucose promoted CCA cell growth and partially rescued cells treated with glucosamine. (B,C) High glucose also rescued the expression of glycoprotein 130 (gp130) and epidermal growth factor receptor (EGFR). (D,E) N -glycosylation in glucosamine-treated CCA cells was significantly increased after high glucose was supplemented, suggesting that increased N -glycosylation is a part of the rescuing mechanisms. Western blots and lectin blots are representative of three biological replications with the same trend of results. Graphs show the averaged band intensities of proteins from three biological replications. GAPDH was used as a loading control, and the control groups (0 mM glucosamine without a 25 mM glucose supplement) were assigned a value of 100%. * p < .05, ** p < .01, *** p < .001.

Article Snippet: Membranes were blocked with 5% skim milk (w/v) in Tris-buffered saline with 0.1% Tween 20 (TBST), and then incubated with primary antibodies included anti-X-linked inhibitor of apoptosis protein (XIAP) (Santa Cruz Biotechnology, Dallas, TX), anti-cyclin D1 (Cell Signaling, Cambridge, MA), anti-p21 (Cell Signaling), anti-caspase 9 (Cell Signaling), anti-caspase 3 (Cell Signaling), anti-cleaved caspase 3 (Cell Signaling), anti-Poly (ADP-ribose) polymerase (PARP) (Cell signaling), anti-cleaved PARP (cell signaling), anti-cyclin dependent kinase (CDK) 4 (Proteintech, Rosemont, IL), anti-CDK6 (Proteintech), anti-epidermal growth factor receptor (EGFR) (Cell Signaling), anti-signal transducer and activator of transcription 3 (STAT3) (Cell Signaling), anti-pSTAT3 (Y075) (Cell Signaling), and anti-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (EMD Millipore, Darmstadt, Germany).

Techniques: Expressing, Glycoproteomics, Western Blot, Control