Fig. 3

LncRNA HOTAIR secreted via exosomes promoted polarization of the M2 phenotype. Exosomes (TU212-Exo and TU177-Exo) were purified for NTA, Electron microscopic and WB analysis. WB analysis for exosomal proteins CD9, CD81 and CD63. NTA analysis of TU212-Exo and TU177-Exo isolated by ultracentrifugation, exosomes nanoparticles relative concentration; Electron microscopic image of TU212-Exo and TU177-Exo. Original gels are presented in Supplementary Fig. 3-A-CD9,Fig. 3-A-CD81 and Fig. 3-A-CD63. B LncRNA HOTAIR was analyzed by performing RT-PCR in TU212 cells or TU177 cells transfected with si-HOTAIR inhibitor, and exosomal lncRNA HOTAIR extracted from TU212-in or TU177-in were also detected. C CD163 and CD206 were detected by using RT-PCR in macrophage treated with TU212-Exo, TU177-Exo, TU212-in-Exo and TU177-in-exo. D, E, F, G, and H Characterization of phenotypes of macrophage by cell surface markers. M2 Macrophages were stained for CD68. Cells were gated for CD68. A representative analysis was presented in TU212-Exo and TU177-Exo, TU212-in-Exo, and TU177-in-Exo. RT-PCR and WB analysis for lncRNA HOTAIR in TU212-Exo and TU177-Exo, TU212-in-Exo, and TU177-in-Exo. Densitometry suggested relative protein expression normalized for GAPDH. Data are shown as mean ± SEM, n = 3 independent experiments. (*p < 0.05, **p < 0.01, *** p < 0.001 were considered statistically significant). Original gels are presented in Supplementary Fig. 3-K-HOTAIR and Fig. 3-K-GAPDH. I CCL18 and IL-10 levers in supernatants of macrophage, measured by ELISA. J and K RT-PCR and WB analysis for lncRNA HOTAIR in TU212-Exo and TU177-Exo, TU212-in-Exo, and TU177-in-Exo