Immunoblotting showing protein expression level with respect to PKCδ and PKCγ.

<p>Western blot analysis showing the expression levels of PKCδ and PKCγ in control, CCI, and ipsilateal SCDH with intramuscular injection with Ad-MOCK or Ad-GDNF (A). The expression levels of PKCδ and PKCγ with respect to each tested group were shown as bar charts of relative ratio normalized with the expression levels of β-actin (B–C). *P<0.05, **P<0.01 compared with the Ad-GDNF group.</p

Immunoblotting showing protein expression level with respect to different NOS isoform.

<p>Western blot analysis showing the expression levels of iNOS, nNOS and eNOS in control, CCI, and ipsilateral SCDH with intramuscular injection with Ad-MOCK or Ad-GDNF (A). The expression levels of iNOS, nNOS and eNOS with respect to each tested group were shown as bar charts of relative ratio normalized with the expression levels of β-actin (B–D). **P<0.01 compared with Ad-GDNF group.</p

Immunoblotting showing protein expression level with respect to IL-6 and IL-1β.

<p>Western blot analysis showing the expression levels of IL-6 and IL-1β in control, CCI, and ipsilateral SCDH with intramuscular injections of Ad-MOCK or Ad-GDNF (A). The expression levels of IL-6 and IL-1β with respect to each tested group were shown as bar charts of relative ratio normalized with the expression levels of β-actin (B–C). *P<0.05, **P<0.01 compared with control group.</p

Immunoblotting showing protein expression level with respect to MMP-2 and MMP-9.

<p>Western blot analysis showing the expression levels of MMP-2 and MMP-9 in control, CCI, and ipsilateral SCDH with intramuscular injection of Ad-MOCK or Ad-GDNF (A). The expression levels of MMP-2 and MMP-9 with respect to each tested group were shown as bar charts of relative ratio normalized with the expression level of β-actin (B–C). *P<0.05 compared with control group.</p

Immunoblotting showing protein expression level with respect to apoptotic and autophagic marker.

<p>Western blot analysis of the effect of CCI on the expression of AIF, caspase-9, cleaved caspase-9, caspase-3, cleaved caspase-3, PARP, cleaved PARP, SPECTRIN, cleaved SPECTRIN, and Beclin-1 on ipsilateral SCDH by intramuscular injection with adenovirus plus GDNF gene (A). Ratios of AIF, cleaved caspase-9, cleaved caspase-3, cleaved PARP, cleaved SPECTRIN, Beclin-1 with β-actin on ipsilateral SCDH were measured using western blot analysis (B–G). *P<0.05, **P<0.01 compared with the CCI group.</p

Immunoblotting showing protein expression level with respect to phosphor-p38, p38 and ED-1.

<p>Western blot analysis showing the expression levels of phospho-p38, p38 and ED-1 in control, CCI, and ipsilateral SCDH with intramuscular injection of Ad-MOCK or Ad-GDNF (A). The expression levels of phospho-p38 and ED-1 with respect to each tested group were shown as bar charts of relative ratio normalized with expression levels of p38 and β-actin, respectively (B-C). *P<0.05, **P<0.01 compared with the Ad-GDNF group. Double immunofluorescence staining of OX42 (D–G), a microglia marker, and phosphor-p38 (H–K) in different tested groups. The expression levels with respect to OX42 and phospho-p38 were obviously enhanced after CCI, but phospho-p38 was no longer highly expressed after administration of Ad-GDNF as shown in merged images (L–O).</p

Double immunofluorescent staining of TUNEL and a neuronal cell marker, NeuN, in the rat ipsilateral SCDH in different treatment groups.

<p>Tissue samples were detected using antibodies against NeuN (A–D) and TUNEL staining for apoptosis (E–H). The merged images show neuron apoptosis in the ipsilateral SCDH (I–L). Yellow arrows indicate TUNEL-positive neurons. The bar chart with respect to fold increase of TUNEL staining positivity (M) and double labeling (TUNEL and NeuN, N) revealed that apoptotic events triggered by CCI were attenuated by Ad-GDNF.</p

Immunoblotting showing protein expression level with respect to GDNF and its receptor.

<p>Western blot analysis showing the expression levels of GDNF and its receptor, GDNFRa-1, under control, CCI, and ipsilateral SCDH with intramuscular injection of Ad-MOCK or Ad-GDNF (A). The expression levels of GDNF and GDNFRa-1 with respect to each tested group were shown as bar charts of relative ratio (B–C). Immunohistochemical (D–K) staining was used to confirm GDNF expression. (D–G: 200X magnification, H–K: 400X magnification) *P<0.05 **P<0.01 compared with control group.</p

The effect of intramuscular delivery of Ad-GDNF on allodynia (A) and thermal hyperalgesia (B) in the CCI model.

<p>*P<0.05 compared with the CCI group at each time point.</p