Journal: Neoplasia (New York, N.Y.)

Article Title: PRAS40 activates the IRE1α-XBP-1-mediated unfolded protein response to exacerbate colorectal cancer by enhancing ST6Gal1-dependent α-2, 6 sialylation of GRP78

doi: 10.1016/j.neo.2026.101297

Figure Lengend Snippet: Effects of ST6Gal1-dependent α-2, 6 sialylation of GRP78 on the UPR. (A) Molecular docking of the interaction between GRP78 and ST6Gal1. GRP78 was shown in blue, and ST6Gal1 was shown in green, respectively. Hydrogen bonds were indicated by yellow dashed lines, and the numbers represented the lengths of hydrogen bonds. (B) Immunofluorescence staining with anti-GRP78 and anti-ST6Gal1 antibodies in HCT116 cells. (C-D) Co-IP followed by western blotting analyses in PRAS40-overexpressed HCT116 cells with anti-ST6Gal1 (C) and anti-GRP78 antibodies (D), respectively. (E-K) The HCT116 cells deleted with ST6Gal1 and overexpressed with Flag-PRAS40, were treated with or without Tg. SNA-pull down assays (E), cell viability analyses (F-G), flow cytometry analyses and the quantification of 3 experiments (H-I), western blotting analyses (J) and PCR analysis (K). Data represent the mean ± SD. Scale bar, 10 μm. ** P < 0.01; *** P < 0.001.

Article Snippet: Antibodies were purchased for detection of PRAS40 (Cell Signaling); GRP78, XBP-1, PARP, IRE1, Flag, GST, ST6Gal1, α-tubulin, β-actin (Proteintech); and SNA (Vector Laboratories).

Techniques: Immunofluorescence, Staining, Co-Immunoprecipitation Assay, Western Blot, Flow Cytometry

Journal: Neoplasia (New York, N.Y.)

Article Title: PRAS40 activates the IRE1α-XBP-1-mediated unfolded protein response to exacerbate colorectal cancer by enhancing ST6Gal1-dependent α-2, 6 sialylation of GRP78

doi: 10.1016/j.neo.2026.101297

Figure Lengend Snippet: Effects of β-sitosterol on ST6Gal1 function and PRAS40-induced cell proliferation. (A) Flow chart for discovery of ST6Gal1 inhibitors. (B) Molecular docking of the interaction of ST6Gal1 protein and β-sitosterol was shown in stick model. ST6Gal1 was shown in blue, β-sitosterol was shown in yellow, and the binding sites of ST6Gal1 were shown in purple. Hydrogen bonds were indicated by yellow dashed lines, and the numbers represented the lengths of hydrogen bonds. (C) Chemical structure of β-sitosterol. (D) SPR assays for the binding affinity of β-sitosterol and ST6Gal1. (E) The ST6Gal1 enzyme activity in the cells treated with or without β-sitosterol. (F-I) The HCT116 cells were transfected with empty vector or ST6Gal1 expression vector followed with or without β-sitosterol treatment (50 μM). SNA-pull down assays (F), representative images (G), cell viability analyses (H), and flow cytometry analyses (I). (J-N) The CRC allografts were established in C57 mice by control or PRAS40-depleted MC38 cells, followed with intraperitoneal injection of β-sitosterol (200 mg/kg), CDDP (3 mg/kg) or combination of both. Experiment flow chart (J), body weight (K), tumor volume (L), tumor weight (M), tumor images (N). Data represent the mean ± SD. Scale bar, 400 μm. * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: Antibodies were purchased for detection of PRAS40 (Cell Signaling); GRP78, XBP-1, PARP, IRE1, Flag, GST, ST6Gal1, α-tubulin, β-actin (Proteintech); and SNA (Vector Laboratories).

Techniques: Binding Assay, Activity Assay, Transfection, Plasmid Preparation, Expressing, Flow Cytometry, Control, Injection