Fig. 1: CRC cell-derived exosomal HSPC111 promotes CAFs activation and correlates with liver metastasis in CRC patients.
A Uptake of DiO-labeled exosomes (green) was analyzed by immunofluorescence microscopy. Scale bar = 20 μm. B Western blot showed HSCs were activated by CRC cell-derived exosomes. C Differentially expressed proteins identified by iTRAQ in LX-2 cells incubated with ExoHCT116. D Bubble diagram of KEGG pathway enrichment analysis related to LX-2 cells incubated with ExoHCT116. E Western blot showed protein level of HSPC111 in different CRC cells and their derived exosomes. F qRT-PCR showed mRNA level of HSPC111 in CAFs have no significant difference compared with control group. G Western blot showed protein level of HSPC111 in different CRC cell-derived exosomes. H Western blot showed exosomal HSPC111 induces the activation of CAFs. I Western blot showed protein level of HSPC111 in serum exosomes isolated from patients and healthy volunteers (n = 6). J Normal fibroblasts (hNFs), non-tumoral fibroblasts (hNTFs) and cancer-associated fibroblasts (hCAFs) isolated from patient liver tissues were photographed by light micrographs (top). Scale bar = 100 μm. Representative immunofluorescence staining of α-SMA and FAP in hNFs, hNTFs and hCAFs isolated from patient liver tissues (bottom). Scale bar = 50 μm. K Western blot showed protein level of HSPC111 in hNFs, hNTFs and hCAFs isolated from patient liver tissues (n = 6). L Scores indicate HSPC111 level in representative CRC tissues. Low group: score 0–1; high group: score 2–3. M Representative images of immunohistochemical staining and quantification of HSPC111expression in NCT, NMCT and CTLM specimens. Scale bar = 50 μm. Each experiment was performed in triplicate. Data are shown as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ns not significant.
