Fig. 3: MLCK210 inhibitor prevents integrin α4 activation. | Nature Communications

Fig. 3: MLCK210 inhibitor prevents integrin α4 activation.

From: PI3Kγ stimulates a high molecular weight form of myosin light chain kinase to promote myeloid cell adhesion and tumor inflammation

Fig. 3

a Quantification of VCAM-Fc binding to IL-1β, SDF-1α, VEGF-A, or Mn2+ stimulated vehicle or MW01-022AZ treated human CD11b+ myeloid cells (n = 3). *p < 0.0001 for IL-1β, SDF-1α, and VEGF-A stimulated, MW01-022AZ vs vehicle-treated cells. No significant difference (ns) for basal (p = 0.9965) or Mn2+ (p = 0.6015) stimulated MW01-022AZ vs vehicle-treated cells. b Flow cytometry profiles of cells from a. c Quantification of HUTS21 or P4C10 antibody binding to basal medium, SDF-1α or Mn2+ stimulated, vehicle or MW01-022AZ treated human CD11b+ cells (n = 3).*p < 0.0001 for HUTS21 binding to SDF-1α stimulated and *p = 0.0002 to Mn2+-stimulated MW01-022AZ vs vehicle-treated cells. No significant difference (ns) for basal MW01-022AZ vs vehicle cells (p > 0.9999). No significant difference (ns) for P4C10 binding to vehicle vs MW01-022AZ treated basal (p = 0.1384), SDF-1α (p > 0.9999) or Mn2+ (p = 0.9997) stimulated cells. d FACs profiles of cells from c. e Quantification of VCAM-Fc binding to SDF-1α or Mn2+ stimulated murine CD11b+ myeloid cells in MW01-022AZ and Vehicle-treated cells (n = 3). *p < 0.0001 for SDF-1α stimulated cells. No significant difference (ns) for basal (p > 0.9999) or Mn2+ (p = 0.9991)-treated cells. f FACs profiles of cells from e. g VCAM-Fc binding (MFI) to basal medium, SDF-1α IL-1β, IL-6, VEGF-A, TNFα, and Mn2+ stimulated murine myeloid cells from WT or Mlck210−/− mice (n = 3). *p < 0.0001 for WT vs Mlck210−/− SDF-1α IL-1β, IL-6, VEGF-A, TNFα-stimulated cells. No significant differences (ns) for secondary antibody alone (p > 0.9999), basal (p = 0.9997), or Mn2+ (p = 0.9928) treated cells. h VCAM-Fc binding (MFI) to basal medium, SDF-1α and Mn2+ stimulated murine myeloid cells from control All stars or Mlck siRNA-transfected cells (n = 3). *p < 0.0001 for SDF1α and Mn2+ stimulated Mlck siRNA vs All Stars transfected cells. No significant differences (ns) for basal Mylk siRNA vs All Stars transfected (p > 0.9999) or untransfected vs All Stars transfected cells (p = 0.9998). For panels a, c, e, g, h, significance determined by one-way Anova with Tukey’s multiple comparisons. Analysis of VCAM-Fc, HUTS, or P4C10 binding was by single color flow cytometry. All replicates are biologically independent samples. Experiments were performed twice. Data are presented as mean values ± SEM. Source data are provided with this article as a Source Data file.

Back to article page