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ROCK inhibition promotes phosphorylation of PI3K/Akt/GSK3β in vivo. (A-E) Increased expression of RhoA, ROCK1/2, <t>GAP43</t> and c-Jun in the LE on day 3 after SNC (n=3). (F-K) Y27632 increases the phosphorylation of PI3K, Akt and GSK3β in the LE on day 3 after SNC (n=3). (L and M) Co-treatment with LY294002 reduces Y27632-induced phosphorylation of GSK3β (n=3). (N) Immunofluorescence images of ROCK1/2 and p-GSK3β in the sciatic nerve and LE. (O-R) Quantification of ROCK1/2 and P-GSK3β expression (n=6). * P<0.05, ** P<0.01 and *** P<0.001. LE, lumbosacral enlargement; SNC, sciatic nerve crush; p-, phosphorylated; NS, not significant (P>0.05).
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ROCK inhibition promotes phosphorylation of PI3K/Akt/GSK3β in vivo. (A-E) Increased expression of RhoA, ROCK1/2, <t>GAP43</t> and c-Jun in the LE on day 3 after SNC (n=3). (F-K) Y27632 increases the phosphorylation of PI3K, Akt and GSK3β in the LE on day 3 after SNC (n=3). (L and M) Co-treatment with LY294002 reduces Y27632-induced phosphorylation of GSK3β (n=3). (N) Immunofluorescence images of ROCK1/2 and p-GSK3β in the sciatic nerve and LE. (O-R) Quantification of ROCK1/2 and P-GSK3β expression (n=6). * P<0.05, ** P<0.01 and *** P<0.001. LE, lumbosacral enlargement; SNC, sciatic nerve crush; p-, phosphorylated; NS, not significant (P>0.05).
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ROCK inhibition promotes phosphorylation of PI3K/Akt/GSK3β in vivo. (A-E) Increased expression of RhoA, ROCK1/2, GAP43 and c-Jun in the LE on day 3 after SNC (n=3). (F-K) Y27632 increases the phosphorylation of PI3K, Akt and GSK3β in the LE on day 3 after SNC (n=3). (L and M) Co-treatment with LY294002 reduces Y27632-induced phosphorylation of GSK3β (n=3). (N) Immunofluorescence images of ROCK1/2 and p-GSK3β in the sciatic nerve and LE. (O-R) Quantification of ROCK1/2 and P-GSK3β expression (n=6). * P<0.05, ** P<0.01 and *** P<0.001. LE, lumbosacral enlargement; SNC, sciatic nerve crush; p-, phosphorylated; NS, not significant (P>0.05).

Journal: International Journal of Molecular Medicine

Article Title: ROCK inhibition promotes axon and myelin regeneration via PI3K/Akt/GSK3β in a mouse sciatic nerve injury model

doi: 10.3892/ijmm.2025.5685

Figure Lengend Snippet: ROCK inhibition promotes phosphorylation of PI3K/Akt/GSK3β in vivo. (A-E) Increased expression of RhoA, ROCK1/2, GAP43 and c-Jun in the LE on day 3 after SNC (n=3). (F-K) Y27632 increases the phosphorylation of PI3K, Akt and GSK3β in the LE on day 3 after SNC (n=3). (L and M) Co-treatment with LY294002 reduces Y27632-induced phosphorylation of GSK3β (n=3). (N) Immunofluorescence images of ROCK1/2 and p-GSK3β in the sciatic nerve and LE. (O-R) Quantification of ROCK1/2 and P-GSK3β expression (n=6). * P<0.05, ** P<0.01 and *** P<0.001. LE, lumbosacral enlargement; SNC, sciatic nerve crush; p-, phosphorylated; NS, not significant (P>0.05).

Article Snippet: GAP43 , Proteintech Group, Inc. , 16971-1-AP , 1:1,000.

Techniques: Inhibition, Phospho-proteomics, In Vivo, Expressing, Immunofluorescence

ROCK inhibition promotes PI3K/Akt/GSK3β phosphorylation and axon regeneration in vitro. (A-a), Customized polydimethylsiloxane mold: The arrowhead and arrow indicate the long and short grooves, respectively. The red spot indicates the site for placement of the DRG, ~1.5 mm away from the intersection. (A-b) Representative image of a DRG placed in the mold. (A-c) Representative image of the DRG immediately after axon transection. A shallow transection mark is left, and the distal stumps of the severed axons are washed away. (A-d) Regenerated axons emerging from the proximal stumps extend across the transection mark without obvious impediment. (B-F) The expression of RhoA, ROCK1/2, GAP43 and c-Jun is significantly elevated after axotomy (n=3). (G-L) Y27632 increases the phosphorylation of PI3K, Akt and GSK3β in the DRG on day 3 after axotomy (n=3). (M and N) LY294002 co-treatment reduces Y27632-induced phosphorylation of GSK3β (n=3). (O and P) The length of regenerated axons is significantly greater in the Y27632 and Y + LY + SB groups (n=5). (Q) Comparison of axon outgrowth in axotomized vs. non-axotomized DRG. *** P<0.001 and **** P<0.0001. DRG, dorsal root ganglion; NS, not significant (P>0.05).

Journal: International Journal of Molecular Medicine

Article Title: ROCK inhibition promotes axon and myelin regeneration via PI3K/Akt/GSK3β in a mouse sciatic nerve injury model

doi: 10.3892/ijmm.2025.5685

Figure Lengend Snippet: ROCK inhibition promotes PI3K/Akt/GSK3β phosphorylation and axon regeneration in vitro. (A-a), Customized polydimethylsiloxane mold: The arrowhead and arrow indicate the long and short grooves, respectively. The red spot indicates the site for placement of the DRG, ~1.5 mm away from the intersection. (A-b) Representative image of a DRG placed in the mold. (A-c) Representative image of the DRG immediately after axon transection. A shallow transection mark is left, and the distal stumps of the severed axons are washed away. (A-d) Regenerated axons emerging from the proximal stumps extend across the transection mark without obvious impediment. (B-F) The expression of RhoA, ROCK1/2, GAP43 and c-Jun is significantly elevated after axotomy (n=3). (G-L) Y27632 increases the phosphorylation of PI3K, Akt and GSK3β in the DRG on day 3 after axotomy (n=3). (M and N) LY294002 co-treatment reduces Y27632-induced phosphorylation of GSK3β (n=3). (O and P) The length of regenerated axons is significantly greater in the Y27632 and Y + LY + SB groups (n=5). (Q) Comparison of axon outgrowth in axotomized vs. non-axotomized DRG. *** P<0.001 and **** P<0.0001. DRG, dorsal root ganglion; NS, not significant (P>0.05).

Article Snippet: GAP43 , Proteintech Group, Inc. , 16971-1-AP , 1:1,000.

Techniques: Inhibition, Phospho-proteomics, In Vitro, Expressing, Comparison

The forward and reverse primers for real-time polymerase chain reaction

Journal: Neural Regeneration Research

Article Title: Nerve root magnetic stimulation regulates the synaptic plasticity of injured spinal cord by ascending sensory pathway

doi: 10.4103/NRR.NRR-D-24-00628

Figure Lengend Snippet: The forward and reverse primers for real-time polymerase chain reaction

Article Snippet: The following antibodies were used: postsynaptic density protein-95 (PSD95; rabbit, 1:1000, Cell Signaling Technology, Danvers, MA, USA, Cat# 3450S, RRID: AB_2292883), Synapsin I (rabbit, 1:1000, Abmart, Shanghai, China, Cat# 334286), GAP43 (mouse, 1:400, Santa Cruz, Biotechnology, Dallas, TX, USA, Cat# sc-17790, RRID: AB_627660), Synapsin II (rabbit, 1:1000, Abclonal, Wuhan, China, Cat# A19542), and GAPDH (mouse, 1:20 000, Sigma-Aldrich, Cat# G8795, RRID: AB_1078991).

Techniques:

NRMS upregulates transcriptional levels of genes related to synaptic plasticity in damaged region of SCI rats. (A) The functional classification of different genes in the SCI + NRMS and SCI + SS groups by GO enrichment analysis. (B) The functional classification of different genes between the SCI + NRMS and SCI + SS groups by KEGG database. (C–F) The relative levels of Syn2 , Syp , GAP43 , and MAP2 mRNA normalized by the Sham group. The bold lines, upper boundaries, and lower boundaries represent the medians, 75 th percentiles, and 25 th percentiles, respectively. Whiskers extend 1.5 times interquartile range ( n = 4 per group). * P < 0.05, vs . Sham group; # P < 0.05, vs . SCI + SS group. GAP43: Growth-associated protein 43; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; MAP2: microtubule-associated protein 2; NRMS: nerve root magnetic stimulation; SCI: spinal cord injury; SS: sham stimulation; Syn2: synapsin II; Syp: synaptophysin.

Journal: Neural Regeneration Research

Article Title: Nerve root magnetic stimulation regulates the synaptic plasticity of injured spinal cord by ascending sensory pathway

doi: 10.4103/NRR.NRR-D-24-00628

Figure Lengend Snippet: NRMS upregulates transcriptional levels of genes related to synaptic plasticity in damaged region of SCI rats. (A) The functional classification of different genes in the SCI + NRMS and SCI + SS groups by GO enrichment analysis. (B) The functional classification of different genes between the SCI + NRMS and SCI + SS groups by KEGG database. (C–F) The relative levels of Syn2 , Syp , GAP43 , and MAP2 mRNA normalized by the Sham group. The bold lines, upper boundaries, and lower boundaries represent the medians, 75 th percentiles, and 25 th percentiles, respectively. Whiskers extend 1.5 times interquartile range ( n = 4 per group). * P < 0.05, vs . Sham group; # P < 0.05, vs . SCI + SS group. GAP43: Growth-associated protein 43; GO: Gene Ontology; KEGG: Kyoto Encyclopedia of Genes and Genomes; MAP2: microtubule-associated protein 2; NRMS: nerve root magnetic stimulation; SCI: spinal cord injury; SS: sham stimulation; Syn2: synapsin II; Syp: synaptophysin.

Article Snippet: The following antibodies were used: postsynaptic density protein-95 (PSD95; rabbit, 1:1000, Cell Signaling Technology, Danvers, MA, USA, Cat# 3450S, RRID: AB_2292883), Synapsin I (rabbit, 1:1000, Abmart, Shanghai, China, Cat# 334286), GAP43 (mouse, 1:400, Santa Cruz, Biotechnology, Dallas, TX, USA, Cat# sc-17790, RRID: AB_627660), Synapsin II (rabbit, 1:1000, Abclonal, Wuhan, China, Cat# A19542), and GAPDH (mouse, 1:20 000, Sigma-Aldrich, Cat# G8795, RRID: AB_1078991).

Techniques: Functional Assay

NRMS alters expression of proteins associated with synaptic plasticity in the injured spinal cord of rats. (A) Representative western blots show the expression levels of PSD95, GAP43, Synapsin I and Synapsin II collected from the damaged area of spinal cord on day 22 after SCI. (B–E) The semi-quantified results of PSD95, GAP43, Synapsin I, and Synapsin II protein levels. All experiments were repeated three times. Data are expressed as mean ± SD ( n = 5 per group). ** P < 0.01, *** P < 0.001, vs. Sham group; # P < 0.05, ## P < 0.01, vs . SCI + SS group. GAP43: Growth-associated protein 43; PSD95: postsynaptic density protein-95; NRMS: nerve root magnetic stimulation; SCI: spinal cord injury; SS: sham stimulation.

Journal: Neural Regeneration Research

Article Title: Nerve root magnetic stimulation regulates the synaptic plasticity of injured spinal cord by ascending sensory pathway

doi: 10.4103/NRR.NRR-D-24-00628

Figure Lengend Snippet: NRMS alters expression of proteins associated with synaptic plasticity in the injured spinal cord of rats. (A) Representative western blots show the expression levels of PSD95, GAP43, Synapsin I and Synapsin II collected from the damaged area of spinal cord on day 22 after SCI. (B–E) The semi-quantified results of PSD95, GAP43, Synapsin I, and Synapsin II protein levels. All experiments were repeated three times. Data are expressed as mean ± SD ( n = 5 per group). ** P < 0.01, *** P < 0.001, vs. Sham group; # P < 0.05, ## P < 0.01, vs . SCI + SS group. GAP43: Growth-associated protein 43; PSD95: postsynaptic density protein-95; NRMS: nerve root magnetic stimulation; SCI: spinal cord injury; SS: sham stimulation.

Article Snippet: The following antibodies were used: postsynaptic density protein-95 (PSD95; rabbit, 1:1000, Cell Signaling Technology, Danvers, MA, USA, Cat# 3450S, RRID: AB_2292883), Synapsin I (rabbit, 1:1000, Abmart, Shanghai, China, Cat# 334286), GAP43 (mouse, 1:400, Santa Cruz, Biotechnology, Dallas, TX, USA, Cat# sc-17790, RRID: AB_627660), Synapsin II (rabbit, 1:1000, Abclonal, Wuhan, China, Cat# A19542), and GAPDH (mouse, 1:20 000, Sigma-Aldrich, Cat# G8795, RRID: AB_1078991).

Techniques: Expressing, Western Blot