Journal: European Journal of Histochemistry : EJH

Article Title: Electroacupuncture combined with rTMS promotes neuronal regeneration via DNMT1-mediated PI3K-AKT pathway in cerebral palsy model

doi: 10.4081/ejh.2026.4533

Figure Lengend Snippet: Effects of electroacupuncture (EA) combined with repetitive transcranial magnetic stimulation (rTMS) on motor function and neuroregeneration-related protein expression in the HIBD-induced cerebral palsy rat model. A ) Behavioral assessments of rotarod latency, grid-walking test, and grip strength test across the Control, HIBD Model, EA, rTMS, and EA+rTMS groups. B ) Western blot analysis of protein expression levels for GAP-43, MBP, PI3K, total AKT, phosphorylated AKT (p-AKT), and DNMT1. C ) Quantitative real-time PCR (qRT-PCR) analysis of mRNA expression levels for GAP-43, MBP, PI3K, total AKT, and DNMT1. Data are presented as mean ± SD (n=6 rats per group); *p <0.05, ** *p <0.01, *** *p <0.001.

Article Snippet: 3) Hypoxia + DNMT1-KD group: following 3 h of hypoxia, neurons were transfected with DNMT1 siRNA (100 nM total concentration, custom sequence; Sangon Biotech, Shanghai, China) using Lipofectamine 3000 (L3000015; ThermoFisher, Waltham, MA, USA) and Opti-MEM (31985070; ThermoFisher) for 6 h, then cultured for an additional 24 h in Neurobasal-A medium under normoxia.

Techniques: Expressing, Control, Western Blot, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: European Journal of Histochemistry : EJH

Article Title: Electroacupuncture combined with rTMS promotes neuronal regeneration via DNMT1-mediated PI3K-AKT pathway in cerebral palsy model

doi: 10.4081/ejh.2026.4533

Figure Lengend Snippet: Electroacupuncture (EA) combined with repetitive transcranial magnetic stimulation (rTMS) enhances neuronal regeneration and modulates DNMT1, GAP-43, and PI3K expression in the rat spinal cord and cerebral cortex. A ) Nissl staining showing neuronal morphology and density in the cerebral cortex across the Control, HIBD Model, EA, rTMS, and EA+rTMS groups, with the EA+rTMS group demonstrating the greatest improvement in neuronal integrity; magnification: 200×. B ) Immunofluorescence staining of DNMT1, GAP-43, and PI3K (with DAPI nuclear counterstain) in the spinal cord and cerebral cortex across all experimental groups; magnification: 200×. C ) Quantitative analysis of relative fluorescence intensity for DNMT1, GAP-43, and PI3K from the immunofluorescence staining assays. Data are presented as mean ±SD (n=6 rats per group); * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: 3) Hypoxia + DNMT1-KD group: following 3 h of hypoxia, neurons were transfected with DNMT1 siRNA (100 nM total concentration, custom sequence; Sangon Biotech, Shanghai, China) using Lipofectamine 3000 (L3000015; ThermoFisher, Waltham, MA, USA) and Opti-MEM (31985070; ThermoFisher) for 6 h, then cultured for an additional 24 h in Neurobasal-A medium under normoxia.

Techniques: Expressing, Staining, Control, Immunofluorescence, Fluorescence

Journal: European Journal of Histochemistry : EJH

Article Title: Electroacupuncture combined with rTMS promotes neuronal regeneration via DNMT1-mediated PI3K-AKT pathway in cerebral palsy model

doi: 10.4081/ejh.2026.4533

Figure Lengend Snippet: DNMT1 knockdown enhances PI3K-AKT signaling and promotes axonal regeneration in hypoxia-exposed primary rat cortical neurons. A ) Western blot analysis of protein expression levels for DNMT1, PI3K, total AKT, phosphorylated AKT (p-AKT), and GAP-43 across the Control, Hypoxia, Hypoxia+DNMT1-KD, Hypoxia+DNMT1-OE, Hypoxia+PI3K Inhibition, and Hypoxia+DNMT1-KD+PI3K-OE groups, with corresponding quantitative analyses below. B ) Quantitative real-time PCR (qRT-PCR) analysis of mRNA expression levels for DNMT1, PI3K, GAP-43, and total AKT in each experimental group. Data are presented as mean ±SD (n=6 independent replicates per group); * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: 3) Hypoxia + DNMT1-KD group: following 3 h of hypoxia, neurons were transfected with DNMT1 siRNA (100 nM total concentration, custom sequence; Sangon Biotech, Shanghai, China) using Lipofectamine 3000 (L3000015; ThermoFisher, Waltham, MA, USA) and Opti-MEM (31985070; ThermoFisher) for 6 h, then cultured for an additional 24 h in Neurobasal-A medium under normoxia.

Techniques: Knockdown, Western Blot, Expressing, Control, Inhibition, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

Journal: European Journal of Histochemistry : EJH

Article Title: Electroacupuncture combined with rTMS promotes neuronal regeneration via DNMT1-mediated PI3K-AKT pathway in cerebral palsy model

doi: 10.4081/ejh.2026.4533

Figure Lengend Snippet: DNMT1 knockdown promotes cell viability and axonal regeneration in hypoxia-exposed primary rat cortical neurons. A ) Cell viability detected by CCK-8 assay across the Control, Hypoxia, Hypoxia+DNMT1-KD, Hypoxia+DNMT1-OE, Hypoxia+PI3K Inhibition, and Hypoxia+DNMT1-KD+PI3K-OE groups. B ) Immunofluorescence staining showing the distribution and expression of DNMT1 (green) and GAP-43 (red), with DAPI (blue) nuclear counterstain and merged images; quantitative analysis of relative fluorescence intensity for DNMT1 and GAP-43 is shown below; magnification: 200×. C ) Axonal length assessment via SAP102 staining (green) with DAPI (blue) nuclear counterstain, and corresponding quantitative analysis of axonal length; magnification: 200×. Data are presented as mean ±SD (n=6 independent replicates per group); * p <0.05, ** p <0.01, *** p <0.001.

Article Snippet: 3) Hypoxia + DNMT1-KD group: following 3 h of hypoxia, neurons were transfected with DNMT1 siRNA (100 nM total concentration, custom sequence; Sangon Biotech, Shanghai, China) using Lipofectamine 3000 (L3000015; ThermoFisher, Waltham, MA, USA) and Opti-MEM (31985070; ThermoFisher) for 6 h, then cultured for an additional 24 h in Neurobasal-A medium under normoxia.

Techniques: Knockdown, CCK-8 Assay, Control, Inhibition, Immunofluorescence, Staining, Expressing, Fluorescence