Fig. 7

Relationship between exosomes and nerve alterations in vivo. A Gross observation of xenograft tumors in nude mice. B Changes in tumor volume were recorded at the time points indicated. C qRT-PCR was used to assess relative expression levels of Gli1 and has-circ-0011536. D HE (200 ×) and immunohistochemical staining (400 ×) of tumors shows the levels of Ki-67. Negative control (NC; tissue without primary antibody) and positive control (PC; human breast carcinoma) was used. E Quantification of Ki-67⁺cells in randomly selected areas of the indicated groups (NC: negative control; PC: positive control). Expression of Ki-67 was quantified as a percentage of Ki-67 positive cells with respect to the total cells from randomly selected areas (n = 5) in each group. Results are means ± SEM. **P < 0.01 for lenti-Gli1 vs. EV-Gli1; ^##P < 0.01 for lenti-Gli1 + si-circ vs. lenti-Gli1. F qRT-PCR was used to assess relative expression levels of mmu-circ-0001261. The correlation between circ-RNA expression and nerve number. G Immunofluorescence staining (40 ×) of tumors indicates the levels of Gli1 and PGP 9.5. Representative Gli1 and PGP9.5 immunostaining in positive control (peripheral nerve sheath tumor specimen). H The number of intra-PDAC nerves was calculated in tissues. *P < 0.05 for lenti-Gli1 vs. EV-Gli1; ^#P < 0.05 for lenti-Gli1 + si-circ vs. lenti-Gli1. I The expression of has-circ-0011536 in exosomes from pancreatic cancer with and without PNR. J Immunofluorescence staining for Gli1, PGP9.5, and VGF on a tissue microarray analysis (TMA) slide containing PDACs. *P < 0.05 for PNR vs. NO PNR