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. 2018 Apr 17;9(29):20640–20657. doi: 10.18632/oncotarget.25045

Figure 4. The CD44 ligand OPN controls invadopodia assembly.

Figure 4

(A) GFP-expressing GSCs were seeded on PLL-coated coverslips for 24 h and stained for cortactin (red). GSCs cultured on PLL for 24 h presented numerous invadopodia puncta. (B) GSC-GFP cells were plated on PLL for 24 h and then stained for secreted OPN ligands (red). Immunofluorescence images revealed OPN localization (red) to regions where GSC-GFP cells adhered. (C) Z-stack analysis confirmed OPN localization to the extracellular space underneath the cells. (D) GSC-GFP spheres were cultured on matrigel-coated coverslips for 2 h, fixed and stained with an OPN antibody (red). Immunofluorescence analysis showed OPN enrichment within the multicellular bulk. (E) GFP-expressing spheres were plated on matrigel-coated substrates for 2 h and stained with a CD44 antibody. OPN deposition within the multicellular bulk was accompanied by CD44 enrichment. Bars: (A) 30 μm; (B) 40 μm; (C) 40 μm; 4 μm; (D) 50 μm; (E) 50 μm.