Journal: International Journal of Oncology
Article Title: Galectin-1 binds GRP78 to promote the proliferation and metastasis of gastric cancer
doi: 10.3892/ijo.2022.5431
Figure Lengend Snippet: GRP78 is associated with Gal-1. (A) The interaction protein network of Gal-1 was revealed by the BioGRID database ( https://thebiogrid.org/ ). (B) GRP78 mRNA expression in GC (n=408) and average (n=211) samples were evaluated using the online tool GEPIA. (C) The expression level of Gal-1 mRNA is independent of GRP78 mRNA in The Cancer Genome Atlas database. (D and E) Gal-1 physically interacted with GRP78. The endogenous proteins from HGC-27 cells were immunoprecipitated with IgG or antibodies against Gal-1 and GRP78, followed by western blot analysis and cell lysis for input. (F) Immunofluorescence colocalization of Gal-1 and GRP78 in GC cells (scale bar, 10 µ m). (G) A western blot assay was performed to detect the GRP78 protein levels in the HGC-27 cells, knocking-down Gal-1. (H) Western blot analysis was performed to detect the GRP78 protein levels in the AGS cells overexpressing Gal-1. * P<0.05. GRP78, glucose-regulated protein 78; Gal-1, galectin-1; GC, gastric cancer; sh-, short hairpin; OE, overexpression.
Article Snippet: The cells were permeabilized with 0.1% Triton X-100 for 15 min. After blocking with 5% bovine serum albumin (Beyotime Institute of Biotechnology) at room temperature for 60 min, the cells were incubated with the following antibodies for 60 min at 37°C: Rabbit anti-human GRP78 (1:300; cat. no. 11587-1-AP) and mouse anti-human Gal-1 (1:100; cat. no. 60223-1-Ig; both from ProteinTech Group, Inc.).
Techniques: Expressing, Immunoprecipitation, Western Blot, Lysis, Immunofluorescence, Over Expression