Journal: Neural Regeneration Research

Article Title: Empowering the NSC-34 cell line as a motor neuron model: Cytosine arabinoside’s action

doi: 10.4103/NRR.NRR-D-24-00034

Figure Lengend Snippet: Neuronal and motor neuronal markers are induced by ara-C. For western blot analysis, NSC-34 cells were plated in growth medium (GM) at 1.5 × 10 5 cells/90 mm dish and protein extracted at 2 hours after plating (T0) or plated at 5 × 10 5 cells/90 mm dish and maintained in differentiation medium supplemented with 0.5 μM ara-C for 4 days (T4), or plated at 1 × 10 6 cells/90 mm dish and maintained in differentiation medium supplemented with 0.5 μM ara-C for 14 days (T14). Proteins were extracted and subjected to SDS-PAGE and western blot. GAP43 (A) and Islet2 (D) analyses represent mean ± SEM of n = 4 experiments, while Vimentin (B) and p75 (C). Bar graphs represent the mean ± SEM of n = 3 experiments. Relative density (A.U., arbitrary units) was expressed as the ratio between the intensity of each target and GAPDH intensity, used as loading control. Statistical analysis was performed by the unpaired t -test. In A, GAP43: T4/T0 * P = 0.03; T14/T0 ** P = 0.002; T14/T4 not significant. In B, Vimentin: T4/T0 not significant; T14/T0 * P = 0.05; T14/T4 not significant. In C, p75: T4/T0 not significant; T14/T0 ** P = 0.006; T14/T4 ** P = 0.006. In D, Islet2: T4/T0 *** P = 0.0007; T14/T0 not significant; T14/T4 **** P = 0.00002. For immunofluorescence analysis, NSC-34 cells were plated at 1 × 10 5 cells/35 mm dish in GM, and analysis performed at 2 hours after plating (T0) or maintained in DM supplemented with 0.5 μM ara-C for 4 (T4) and 14 (T14) days. Representative images of immunofluorescence confocal microscopy at different time points are shown. Green staining was used to visualize GAP43 (Panel A) and p75 (Panel C) markers, while red staining was used for Vimentin (Panel B) and Islet 2 (Panel D). Scale bars: 50 or 100 μm, as indicated. ara-C: Cytosine arabinoside; GAP43: growth associated protein 43; GAPDH: glyceraldehyde 3-phosphate dehydrogenase.

Article Snippet: Mouse, GAP43 , 1:1000 , 1:1000 , MyBioSource, San Diego, CA, USA , MBS555937.

Techniques: Western Blot, SDS Page, Control, Immunofluorescence, Confocal Microscopy, Staining

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

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

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:

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

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