Journal: Molecular medicine reports

Article Title: CXCL12/CXCR4 signaling pathway regulates cochlear development in neonatal mice.

doi: 10.3892/mmr.2016.5085

Figure Lengend Snippet: Figure 1. Immunofluorescence of CXCL12 and CXCR4 in spiral ganglion neurons of neonatal mouse inner ear. CXCL12 and CXCR4 expression in spiral ganglion neurons were detected by immunostaining at (A) P0, (B) P7, (C) P14 and (D) P21. Nuclei were stained with DAPI and merged images of red, green and DAPI staining are shown. Images were captured under a fluorescence microscope (magnification, x40). CXCL12, CXC chemokine ligand 12; CXCR4, CXC chemokine receptor 4; DAPI, 4'‑6‑diamidino‑2‑phenylindole dihydrochloride; P, postnatal day.

Article Snippet: The protein extract was centrifuged and the concentration of CXCL12 in the supernatant was determined using a commercially available CXCL12 ELISA kit (cat. no. CSB-E04723m; Cusabio Biotech Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques: Immunofluorescence, Expressing, Immunostaining, Staining, Fluorescence, Microscopy

Journal: Molecular medicine reports

Article Title: CXCL12/CXCR4 signaling pathway regulates cochlear development in neonatal mice.

doi: 10.3892/mmr.2016.5085

Figure Lengend Snippet: Figure 2. Quantitative analysis of relative fluorescence intensity. Relative fluorescence intensity of (A) CXCL12 and (B) CXCR4 in the different groups, fol lowing normalization with the P0 group. Fluorescence intensity was quantified using ImageJ software. *P<0.05 and **P<0.01 vs. P0; &P<0.05 vs. P7. Data are presented as the mean ± standard deviation. CXCL12, CXC chemokine ligand 12; CXCR4, CXC chemokine receptor 4; P, postnatal day.

Article Snippet: The protein extract was centrifuged and the concentration of CXCL12 in the supernatant was determined using a commercially available CXCL12 ELISA kit (cat. no. CSB-E04723m; Cusabio Biotech Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques: Fluorescence, Software, Standard Deviation

Journal: Molecular medicine reports

Article Title: CXCL12/CXCR4 signaling pathway regulates cochlear development in neonatal mice.

doi: 10.3892/mmr.2016.5085

Figure Lengend Snippet: Figure 3. mRNA and protein expression levels of CXCL12 and CXCR4 in spiral ganglion neurons of neonatal mice. (A) Relative expression levels of CXCL12 and CXCR4 mRNA on P0, P7, P14 and P21 detected by reverse transcription‑quantitative polymerase chain reaction analysis. (B) Concentration of CXCL12 protein was quantified by enzyme‑linked immunosorbent assay in the different groups. (C) Protein expression of CXCR4 in spiral ganglion neurons of neonatal mice detected by western blot analysis. (D) Relative protein expression was quantified using Image J software. *P<0.05 and **P<0.01 vs. P0; &P<0.05 vs. P7. Data are presented as the mean ± standard deviation. CXCL12, CXC chemokine ligand 12; CXCR4, CXC chemokine receptor 4; P, postnatal day.

Article Snippet: The protein extract was centrifuged and the concentration of CXCL12 in the supernatant was determined using a commercially available CXCL12 ELISA kit (cat. no. CSB-E04723m; Cusabio Biotech Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques: Expressing, Polymerase Chain Reaction, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Software, Standard Deviation

Journal: Molecular medicine reports

Article Title: CXCL12/CXCR4 signaling pathway regulates cochlear development in neonatal mice.

doi: 10.3892/mmr.2016.5085

Figure Lengend Snippet: Figure 4. CXCL12 enhances the expression of CXCR4 in spiral ganglion neurons. (A) Reverse transcription‑quantitative polymerase chain reac tion analysis of CXCR4 mRNA expression in spiral ganglion neurons. Spiral ganglion neurons were treated with 100 ng/ml CXCL12, with or without 20 µg/ml AMD3100 for 7 days. *P<0.05 vs. control group; &P<0.05 vs. CXCL12‑treated group. (B) Western blot analysis of CXCR4 protein expression in different treatment groups. (C) Relative protein expression level of CXCR4 was quantified using Image J software. *P<0.05 vs. con trol group; &&P<0.01 vs. CXCL12‑treated group. Data are presented as the mean ± standard deviation. CXCL12, CXC chemokine ligand 12; CXCR4, CXC chemokine receptor 4.

Article Snippet: The protein extract was centrifuged and the concentration of CXCL12 in the supernatant was determined using a commercially available CXCL12 ELISA kit (cat. no. CSB-E04723m; Cusabio Biotech Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques: Expressing, Control, Western Blot, Software, Standard Deviation

Journal: Molecular medicine reports

Article Title: CXCL12/CXCR4 signaling pathway regulates cochlear development in neonatal mice.

doi: 10.3892/mmr.2016.5085

Figure Lengend Snippet: Figure 5. CXCL12/CXCR4 signaling inhibits the apoptosis of spiral ganglion neurons. (A) Western blot analysis of caspase‑3 and cleaved caspase‑3 protein expression in spiral ganglion neurons. Spiral ganglion neurons were treated with 100 ng/ml CXCL12 with or without 20 µg/ml AMD3100 for 7 days. (B) Relative protein expression levels of caspase‑3 and cleaved caspase‑3 were quantified using Image J software. *P<0.05 vs. control, day 7; &P<0.05 vs. CXCL12‑treated group, day 7. Data are presented as the mean ± standard deviation. CXCL12, CXC chemokine ligand 12; CXCR4, CXC chemokine receptor 4.

Article Snippet: The protein extract was centrifuged and the concentration of CXCL12 in the supernatant was determined using a commercially available CXCL12 ELISA kit (cat. no. CSB-E04723m; Cusabio Biotech Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques: Western Blot, Expressing, Software, Control, Standard Deviation

Journal: Molecular medicine reports

Article Title: CXCL12/CXCR4 signaling pathway regulates cochlear development in neonatal mice.

doi: 10.3892/mmr.2016.5085

Figure Lengend Snippet: Figure 7. Photomicrographs illustrating spiral ganglion neurons (magnification, x40). Spiral ganglion neurons were stained by neuron‑specific enolase antibody in the (A) control and (B) CXCL12/CXCR4‑blocked groups. The mice were administered with CXCR4 antagonist, AMD3100, by intraperitoneal injection for 48 h. The spiral ganglion was then isolated and detected by immunohistochemistry using neuron‑specific enolase antibody. The spiral ganglion neurons exhibited a normal morphology in the control group, but were reduced in number and had a fuzzy cell morphology in AMD3100‑treated cells. Mice treated with phosphate‑buffered saline served as the control. CXCL12, CXC chemokine ligand 12; CXCR4, CXC chemokine receptor 4.

Article Snippet: The protein extract was centrifuged and the concentration of CXCL12 in the supernatant was determined using a commercially available CXCL12 ELISA kit (cat. no. CSB-E04723m; Cusabio Biotech Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques: Staining, Control, Injection, Isolation, Immunohistochemistry, Saline

Journal: Molecular medicine reports

Article Title: CXCL12/CXCR4 signaling pathway regulates cochlear development in neonatal mice.

doi: 10.3892/mmr.2016.5085

Figure Lengend Snippet: Figure 6. CXCL12/CXCR4 regulates the growth of dendrites in spiral ganglion neurons in vitro. (A) Western blot analysis of MAP2 protein expression in spiral ganglion neurons treated with CXCL12 in the presence or absence of 20 µg/ml AMD3100 for 7 days. (B) Relative protein expression level of MAP2 was quantified using Image J software. *P<0.05 vs. control, day 7; &P<0.05 vs. CXCL12‑treated group, day 7. (C) The mean number of dendrites per cell in the various treated groups. The mean number of dendrites were calculated on day 7. *P<0.05 vs. control; &P<0.05 vs. CXCL12‑treated group. (D) The mean length of dendrites per cell in the various treatment groups. *P<0.05 vs. control; &P<0.05 vs. CXCL12‑treated group. Data are presented as the mean ± standard deviation. MAP2, microtubule‑associated protein 2; CXCL12, CXC chemokine ligand 12; CXCR4, CXC chemokine receptor 4.

Article Snippet: The protein extract was centrifuged and the concentration of CXCL12 in the supernatant was determined using a commercially available CXCL12 ELISA kit (cat. no. CSB-E04723m; Cusabio Biotech Co., Ltd., Wuhan, China) according to the manufacturer's protocol.

Techniques: In Vitro, Western Blot, Expressing, Software, Control, Standard Deviation