rat schwann cells  (ATCC)

Journal: Biomolecules & Therapeutics

Article Title: Clemastine Restores Myelination Protein Expression in S16 Schwann Cells by Enhancing AMPK Activation and Reducing H 2 O 2 -Induced Oxidative Stress

doi: 10.4062/biomolther.2025.188

Figure Lengend Snippet: Determination of optimal Clemastine concentration and time-dependent activation of AMPK signaling in S16 Schwann cells. (A) S16 cells were treated with various concentrations of Clemastine (0.1, 0.5, 1, 5, and 10 μM) for 24 h, and cell viability was assessed using the CCK-8 assay. Cell viability is presented as mean ± SD from three independent experiments. Statistical analysis was performed using Student’s t-test. *** p <0.0005 vs. Vehicle group. (B) Representative microscopic images of S16 Schwann cells treated with increasing concentrations of Clemastine (0.5, 1, 5 and 10 μM) for 24 h. Images were taken under a phase-contrast microscope at 200× magnification. (C) S16 cells were treated with Clemastine (1 μM) for 0, 24, and 48 h, and protein levels of pAMPK(T172), total AMPK, and MPZ were analyzed by Western blotting. (D) Quantitative analysis of Western blot bands confirmed the time- dependent increase of AMPK activation and MPZ expression following Clemastine treatment. Results are expressed as mean ± SD from three independent experiments. Statistical significance was determined with alphabetical indication using one-way ANOVA followed by Tukey’s test versus each group ( p <0.05). Statistical significance is indicated as follows: * p <0.05, ** p <0.01, and *** p <0.001, ns; not significant.

Article Snippet: Immortalized rat Schwann cells (S16) were purchased from ATCC (Bethesda, MD, USA).

Techniques: Concentration Assay, Activation Assay, CCK-8 Assay, Microscopy, Western Blot, Expressing

Journal: Biomolecules & Therapeutics

Article Title: Clemastine Restores Myelination Protein Expression in S16 Schwann Cells by Enhancing AMPK Activation and Reducing H 2 O 2 -Induced Oxidative Stress

doi: 10.4062/biomolther.2025.188

Figure Lengend Snippet: Immunofluorescence analysis of Clemastine-induced AMPK activation and myelin-related protein expression in S16 Schwann cells. Immunofluorescence staining of S16 cells after 48 h Clemastine treatment showed increased immunoreactivity for (A) pAMPK(T172) and (B) MPZ. Images were captured using a fluorescence microscope at 400× magnification.

Article Snippet: Immortalized rat Schwann cells (S16) were purchased from ATCC (Bethesda, MD, USA).

Techniques: Immunofluorescence, Activation Assay, Expressing, Staining, Fluorescence, Microscopy

Journal: Biomolecules & Therapeutics

Article Title: Clemastine Restores Myelination Protein Expression in S16 Schwann Cells by Enhancing AMPK Activation and Reducing H 2 O 2 -Induced Oxidative Stress

doi: 10.4062/biomolther.2025.188

Figure Lengend Snippet: Establishment of a H 2 O 2 -induced oxidative stress model and its inhibitory effect on Clemastine-induced AMPK and MPZ expression in S16 Schwann cells. (A) S16 cells were exposed to increasing concentrations of hydrogen peroxide (H 2 O 2 ) 0, 0.1, 1, 5, 10, and 50 μM for 1 h, followed by a 24-h incubation in fresh medium. Cell viability was measured using the CCK-8 assay and presented as mean ± SD from three independent experiments. (B) Representative phase-contrast microscopic images of S16 cells following H 2 O 2 exposure. Cells treated with ≥10 μM H 2 O 2 exhibited clear morphological changes, including shrinkage, membrane blebbing, rounding, and detachment, consistent with early cytotoxic stress responses. (C) Western blot analysis of S16 cells treated with Clemastine (1 µM) with H 2 O 2 (5 µM). Expression levels of pAMPK(T172), total AMPK, and MPZ were evaluated to assess the impact of oxidative stress on Clemastine-induced signaling. (D) Densitometric quantification of protein bands normalized to β-Actin. Statistical significance was determined using Student’s t-test vs Clemastine only from Cle/H 2 O 2 co-treated samples. Statistical significance is indicated as follows: * p <0.05, ** p <0.01, and *** p <0.001, ns; not significant.

Article Snippet: Immortalized rat Schwann cells (S16) were purchased from ATCC (Bethesda, MD, USA).

Techniques: Expressing, Incubation, CCK-8 Assay, Membrane, Western Blot

Journal: Biomolecules & Therapeutics

Article Title: Clemastine Restores Myelination Protein Expression in S16 Schwann Cells by Enhancing AMPK Activation and Reducing H 2 O 2 -Induced Oxidative Stress

doi: 10.4062/biomolther.2025.188

Figure Lengend Snippet: Metformin restores Clemastine-induced pAMPK and myelin protein expression at the cellular level under H 2 O 2 -induced oxidative stress. Representative immunofluorescence images of S16 Schwann cells treated with Clemastine (1 µM) and hydrogen peroxide (H 2 O 2 , 5 µM), with or without Metformin (Met, 2 mM) for 24 h. Staining for (A) pAMPK(T172) and (B) MPZ demonstrated that co-treatment with H 2 O 2 reduced the Clemastine-induced signal intensity. Met supplementation restored expression levels of both proteins, indicating the recovery of myelin-related signaling. Images were acquired at 400× magnification.

Article Snippet: Immortalized rat Schwann cells (S16) were purchased from ATCC (Bethesda, MD, USA).

Techniques: Expressing, Immunofluorescence, Staining

Journal: Biomolecules & Therapeutics

Article Title: Clemastine Restores Myelination Protein Expression in S16 Schwann Cells by Enhancing AMPK Activation and Reducing H 2 O 2 -Induced Oxidative Stress

doi: 10.4062/biomolther.2025.188

Figure Lengend Snippet: Enhanced AMPK activation by Metformin inhibits ROS and lipid accumulation from Clemastine/H 2 O 2 co-treated S16 Schwann cell. Metformin (2 mM) treatment in S16 Schwann cells exposed to Clemastine (1 µM) and hydrogen peroxide (H 2 O 2 , 5 µM) resulted in (A) reduced intracellular ROS levels, as determined by DCFDA fluorescence staining. (B) Oil Red O staining demonstrated increased intracellular lipid accumulation (lipid droplet deposition) following Clemastine/H 2 O 2 co-treatment, which was significantly attenuated by Met addition. (C) Western blot analysis of lipid metabolism- related proteins revealed that Clemastine/H 2 O 2 co-treatment modestly increased PPARα expression while elevating FASN levels. Met treatment recovered both PPARα and MPZ but reduced FASN expression. Statistical significance is indicated as follows: *** p <0.001, ns; not significant.

Article Snippet: Immortalized rat Schwann cells (S16) were purchased from ATCC (Bethesda, MD, USA).

Techniques: Activation Assay, Fluorescence, Staining, Western Blot, Expressing