Fig 1: M6PR and EphB4 are enriched in exosomes from SRGN-overexpressing ESCC cells. (A) GO analysis of differentially expressed proteins in exosomes from KYSE150-SRGN. (B) PANTHER™ Pathway enrichment analysis for differentially expressed proteins in exosomes from KYSE150-SRGN. All enriched pathways were shown. (C) Western blot validation of the upregulated proteins identified by LC-MS/MS in SRGN Exo compared with Con Exo. The concentration of exosomes was measured by Nanosight NS500 and equal numbers of exosomes were used for Western blot analysis. (D) Western blot analysis of M6PR, EphB4, ALIX and CD63 expressions in density gradient fractions of exosomes from ESCC cells. After flotation of crude exosomes in iodixanol gradients, equal volumes of each fraction were used for Western blot analysis.
Fig 2: Exosomal M6PR mediates the effect of SRGN on angiogenesis in vitro and in vivo. (A) Effect of M6PR Exo on tube formation ability of HUVECs. Scale bar, 200 µm. Data are presented as mean ± SD. n = 7. ***, P < 0.001. (B) Western blot analysis of M6PR in cell lysates, CM and exosomes of ESCC cells with manipulated SRGN and M6PR expression. (C) Representative images of HUVECs treated with indicated CMs from ESCC cells. Scale bar, 200 µm. (D) Quantification of the numbers of nodes, junctions, segments and meshes in (C). Data are presented as mean ± SD. n = 8 and 3 for KYSE150 and KYSE410, respectively. *, P < 0.05; ***, P < 0.001. (E) Representative images of HUVECs treated with indicated Exo from ESCC cells. Scale bar, 200 µm. (F) Quantification of the numbers of nodes, junctions, segments and meshes in (E). Data are presented as mean ± SD. n = 6. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (G) Effects of exosomes from ESCC cells with SRGN overexpression and M6PR-knockdown and rhM6PR on in vivo angiogenesis. Data are presented as mean ± SD. n = 4. *, P < 0.05.
Fig 3: M6PR has prognostic significance in ESCC patients. (A) Correlation analysis between serum SRGN and M6PR in 98 patients with ESCC. (B) Kaplan-Meier curves comparing the survival outcome of ESCC patients with high versus low serum M6PR expression.
Supplier Page from OriGene Technologies for M6PR (NM_002355) Human Recombinant Protein