Figure 5.

Impact of ASML^wt and ASML-CD44v^kd exosomes on mRNA and protein expression in LnStr and LuFb. (A–D) LnStr and LuFb were co-cultured for 48 hours with ASML^wt and ASML-CD44v^kd exosomes. (A and B) Cells were harvested, washed, and lysed and mRNA was extracted and analyzed (Ilumina and SurePrintG3Rat-GE-8x60K microarray). (A) Number of LnStr mRNA with a more than two-fold change after co-culture with ASML^wt or ASML-CD44v^kd exosomes. (B) mRNA in LnStr that are more than two-fold upregulated after co-culture with exosomes and comparison of the relative mRNA amount in exosomes versus LnStr. (A and B) Mean values of duplicates, respectively, triplicates, of two independent microarray analyses. (B) Significant differences between LnStr and LnStr co-cultured with ASML exosomes: *. (C) Upregulated gene expression (selected examples) in LnStr co-cultured with exosomes was confirmed by qRT-PCR (mean of three replicate samples with SD < 0.25, indicating reliability according to the software program for ΔC_T) and (D) in LnStr and LuFb at the protein level by flow cytometry. The mean percentage of stained cells (triplicates) are shown; significant differences between untreated LnStr/LuFb and LnStr/LuFb co-cultured with ASML^wt exosomes: black *; significant differences between untreated LnStr/LuFb and LnStr/LuFb co-cultured with ASML-CD44v^kd exosomes: gray *. (C and D) Experiments were repeated three times revealing comparable results. Abbreviations: MMP3, matrix metalloproteinase 3; CXCL2, chemokine ligand 2; CCL20, chemokine ligand 20; MT1a, metallothionein; PTGS2, prostaglandin-endoperoxide synthase 2; alpha2M, alpha-2-macroglobulin; RGS2, regulator of G-protein signaling 2; PRG4, proteoglycan 4; VCAM1, vascular cell adhesion molecule-1/CD106; ANKRD1, ankyrin repeat domain 1; PLA2g2A, phospholipase A2 group 2A; SOD2, superoxide dismutase 2; CCL19, chemokine ligand 19; SLC40A1, solute carrier family 39; GADD45g, growth arrest and DNA-damage-inducible 45γ; SLPI, secretory leukocyte peptidase inhibitor; BTG2, B-cell translocation gene 2; ICAM1, intercellular adhesion molecule 1/CD54; MYH11, myosin heavy chain 11; AMACR, α-methylacyl-CoA racemase; MPG, matrix Gla protein; ADAMTS8, a disintegrin-like and metalloprotease with thrombospondin type 1, motif 8; RASl11b, RAS-like family 11 member B; EDN1, endothelin 1; SSB-1, SPRY domain-containing SOCS box protein SSB-1; FABP3, fatty acid binding protein 3; FST, follistatin; ADAMTS5, ADAMTS, motif 5; CXCL1, chemokine ligand 1; SCBG1A1, secretoglobin, family 1A, member 1; PPP1R3c, protein phosphatase 1, regulatory subunit 3C; CD49a, integrin α1; MDR1, ATP-binding cassette, subfamily B, member 1; KRT2-7, keratin complex 2; NBI1, neuroblastoma suppression of tumorigenicity; latent TGFβ, latent TGFβ binding protein.