Fig. 4
Exosomal CD44 is a functional mediator in intercellular communication. a Real-time qRT-PCR and Western blotting. qRT-PCR of CD44 mRNA expression after transfecting with CD44 overexpression plasmid for 24 h and treating with 500μg/ml of G418 for 2 weeks. Cell transfected with the empty vectors acted as controls. Results were shown as means ± SEM for three separate experiments, ** p < 0.01. Western blot analysis on cells mentioned above to validate the protein level of CD44 (10 μg/lane). The blots shown were representative of three separate experiments. b Cell proliferation assay. 4 × 103 HO8910 cells transfected with CD44 overexpression plasmids and with empty vectors were added to 96-well plates. 24 h, 48 h, 72 h, 96 h of cell viability were respectively assessed by CCK8 assays. Results in B were shown as means ± SEM for three separate experiments,*** p < 0.001,****p < 0.0001. c-h migration and invasion assay. 2 × 105 HO8910 cells transfected with CD44 overexpression plasmids and with empty vectors were respectively added to trans-well plates, and 3 × 105 cells mentioned above were added to with trans-well plates with matrigel-coated.24 h later, migration and invasion were assessed by trans-well assays. Results in C and F were representative of three experiments. Results in D, E, G, H were shown as means ± SEM for three separate experiments, **p < 0.01, *** p < 0.001, **** p < 0.0001