Journal: bioRxiv
Article Title: Galectin-1 Modulates Cell Adhesions, Caveolae, and Vascular Permeability in Kidney Endothelial Cells – Insights from Proteomics, Phosphoproteomics, and Functional Studies
doi: 10.64898/2026.03.03.709385
Figure Lengend Snippet: A) Overview of treatment and phosphoproteome enrichment workflow. B) Phosphopeptides identified, quantified, and significant for each treatment and their interaction after correcting for matched unmodified peptides using MSStatsPTM in R. C) Volcano plots of phosphopeptide Log2FC of IFNγ treatment, LGALS1 siRNA, and both IFNγ and LGALS1 siRNA, compared to non-targeting control and vehicle plotted against –log10(p-value). Orange colour represents non-significant proteins, while blue colour represents those with p<0.01 (linear mixed-effects model), and green represents those with FDR: BH<0.2. D) Heatmap of 28 phosphopeptides significant in at least 1 comparison from panel C. Red colour indicates increased expression, blue colour indicates decreased expression, and black colour indicates phosphopeptides that were not found in that comparison. Colour on the top bar denotes function (obtained from uniport.org ) of the protein the phosphopeptide belongs to. Black and grey asterisks denote FDR<0.2 and p<0.01, respectively. Immunostaining of CAVN1 and CAV1 when GMECs were treated with LGALS1 siRNA, IFNγ, or both. Green colour indicates CAV1, red colour indicates CAVN1. Red arrows denote co-staining of CAV1 and CAVN1 in the NT_V condition. LGALS1 , galectin-1 gene; GMEC, glomerular microvascular endothelial cells; IFNγ, interferon-γ; NT, non-targeting control; NT_V, non-targeting control and vehicle; PTM, post-translational modification.
Article Snippet: Samples were incubated overnight at 4°C with rabbit polyclonal anti-galectin-1 antibody (1:100; GTX101566; GeneTex), rabbit polyclonal anti-CAVN1 antibody (1:1000; 18892-1-AP; Proteintech), or mouse monoclonal anti-CAV1 antibody (1:1000; 66067-1-IG; Proteintech).
Techniques: Phospho-proteomics, Control, Comparison, Expressing, Immunostaining, Staining, Modification