Journal: International Journal of Molecular Medicine

Article Title: Pericyte-derived extracellular vesicles-mimetic nanovesicles improves peripheral nerve regeneration in mouse models of sciatic nerve transection

doi: 10.3892/ijmm.2021.5073

Figure Lengend Snippet: PC-NVs induce neurovascular regeneration by enhancing NF expression and survival signaling in SNT model mice. (A) Representative western blot for NFs (BDNF, NT-3 and NGF), survival signaling (activation of PI3K and Akt) and cell death signaling (activation of JNK and c-Jun) in sciatic nerve tissue from SNT model mice at 5 days after injection with HBS or PC-NVs (1 and 5 µ g). Data are presented as the relative density of (B) BDNF, (C) NGF, (D) NT-3, (E) p-PI3K/PI3K, (F) p-Akt/Akt, (G) p-JNK/JNK and (H) p-c-Jun/c-Jun, with β-actin as the loading control. Each bar depicts the mean ± SEM of four repeats. The relative ratio of the HBS group was arbitrarily set to 1. * P<0.05 vs. HBS group; # P<0.05 vs. PC-NVs (1 µ g) group. ns, not significant. HBS, HEPES buffer solution; PC-NV, pericyte-derived extracellular vesicle-mimetic nanovesicle; SNT, sciatic nerve transection; NF, neurotrophic factor; BDNF, brain-derived nerve growth factor; NT-3, neurotrophin-3; NGF, nerve growth factor.

Article Snippet: After blocking with 5% non-fat dry milk for 1.5 h at room temperature, the membranes were incubated at room temperature with antibodies against brain-derived nerve growth factor (BDNF; 1:500; cat. no. sc-546; Santa Cruz Biotechnology, Inc.), nerve growth factor (NGF; 1:500; cat. no. sc-548; Santa Cruz Biotechnology, Inc.), neurotrophin-3 (NT-3; 1:500; cat. no. sc-547; Santa Cruz Biotechnology, Inc.), phosphorylated (p)-PI3K (1:500; cat. no. 4228; Cell Signaling Technology, Inc.), total (t)-PI3K (1:1,000; cat. no. 4292; Cell Signaling Technology, Inc.), p-Akt (1:500; cat. no 9271; Cell Signaling Technology, Inc.), t-Akt (1:1,000; cat. no. 9272; Cell Signaling Technology, Inc.), p-c-JUN (1:500; cat. no. 9261; Cell Signaling Technology, Inc.), t-c-JUN (1:1,000; cat. no. 9165; Cell Signaling Technology, Inc.), p-SAPK/JNK (1:500; cat. no. 9251; Cell Signaling Technology, Inc.), t-JNK (1:1,000; cat. no. 9252; Cell Signaling Technology, Inc.) and β-actin (1:2,000; cat. no. ab16051; Abcam) for 2 h. The membranes were washed three times for 10 min with PBST (0.1% Tween-20) at room temperature.

Techniques: Expressing, Western Blot, Activation Assay, Injection, Control, Derivative Assay