nsc-34 mouse neuron cell line  (Procell Inc)

Journal: Regenerative Therapy

Article Title: Tanshinone IIA protects motor neuron-like NSC-34 cells against lipopolysaccharide-induced cell injury by the regulation of the lncRNA TCTN2/miR-125a-5p/DUSP1 axis

doi: 10.1016/j.reth.2023.03.007

Figure Lengend Snippet: LPS induces cell injury and inhibits TCTN2 expression in NSC-34 cells. NSC-34 cells were treated with LPS for 24 h at various concentrations of 0, 25, 50, 75 and 100 ng/mL. (A) Cell viability by CCK-8 assay. (B) Representative images depicting a cell apoptosis assay and cell apoptosis by flow cytometry. (C) The levels of TNF-α, IL-6 and IL-1β by ELISA assay using the assay kits. (D) qRT-PCR for relative TCTN2 expression in treated cells. LPS: lipopolysaccharide. ∗ P < 0.05.

Article Snippet: The mouse motor neuron-like NSC-34 cell line was provided by Cedarlane (Burlington, NC, USA) and cultivated at 37 °C, 5% CO 2 in Dulbecco's modified Eagle Medium (Life Technologies, Scotland, UK) plus 1% penicillin/streptomycin (Life Technologies) and 10% fetal calf serum (Biosera, Boussens, France).

Techniques: Expressing, CCK-8 Assay, Apoptosis Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Regenerative Therapy

Article Title: Tanshinone IIA protects motor neuron-like NSC-34 cells against lipopolysaccharide-induced cell injury by the regulation of the lncRNA TCTN2/miR-125a-5p/DUSP1 axis

doi: 10.1016/j.reth.2023.03.007

Figure Lengend Snippet: TSIIA alleviates cell damage induced by LPS in NSC-34 cells. NSC-34 cells were simultaneously treated with 100 ng/mL of LPS and the indicated doses (0, 2, 4, 6 and 8 μM) of TSIIA for 24 h. (A) CCK-8 assay for the viability of cells after various treatments. (B) Representative images showing a cell apoptosis assay and cell apoptosis by flow cytometry. (C) ELISA assay for TNF-α, IL-6 and IL-1β levels using the corresponding assay kit. (D) Relative expression of TCTN2 by qRT-PCR in treated NSC-34 cells. LPS: lipopolysaccharide, TSIIA: Tanshinone IIA.∗ P < 0.05.

Article Snippet: The mouse motor neuron-like NSC-34 cell line was provided by Cedarlane (Burlington, NC, USA) and cultivated at 37 °C, 5% CO 2 in Dulbecco's modified Eagle Medium (Life Technologies, Scotland, UK) plus 1% penicillin/streptomycin (Life Technologies) and 10% fetal calf serum (Biosera, Boussens, France).

Techniques: CCK-8 Assay, Apoptosis Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR

Journal: Regenerative Therapy

Article Title: Tanshinone IIA protects motor neuron-like NSC-34 cells against lipopolysaccharide-induced cell injury by the regulation of the lncRNA TCTN2/miR-125a-5p/DUSP1 axis

doi: 10.1016/j.reth.2023.03.007

Figure Lengend Snippet: TSIIA alleviates LPS-induced cell injury by elevating TCTN2 expression. NSC-34 cells were transfected with or without si-NC or si-TCTN2 and then treated with or without LPS (100 ng/mL) and TSIIA (0 or 8 μM). (A) TCTN2 expression by qRT-PCR in transfected cells. (B) Cell viability by CCK-8 assay. (C) Representative images showing a cell apoptosis assay and cell apoptosis by flow cytometry. (D) ELISA assay for TNF-α, IL-6 and IL-1β levels using the corresponding assay kit. (E) Representative images depicting a western blot assay and the levels of PCNA and Bcl-2 by western blot. (F) SOD activity using the assay kit. (G) MDA level using the assay kit. LPS: lipopolysaccharide, TSIIA: Tanshinone IIA. ∗ P < 0.05.

Article Snippet: The mouse motor neuron-like NSC-34 cell line was provided by Cedarlane (Burlington, NC, USA) and cultivated at 37 °C, 5% CO 2 in Dulbecco's modified Eagle Medium (Life Technologies, Scotland, UK) plus 1% penicillin/streptomycin (Life Technologies) and 10% fetal calf serum (Biosera, Boussens, France).

Techniques: Expressing, Transfection, Quantitative RT-PCR, CCK-8 Assay, Apoptosis Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Activity Assay

Journal: Regenerative Therapy

Article Title: Tanshinone IIA protects motor neuron-like NSC-34 cells against lipopolysaccharide-induced cell injury by the regulation of the lncRNA TCTN2/miR-125a-5p/DUSP1 axis

doi: 10.1016/j.reth.2023.03.007

Figure Lengend Snippet: TCTN2 directly targets miR-125a-5p. (A) Sequence of miR-125a-5p, the binding sites for miR-125a-5p in TCTN2 and the mutation of the target sites. (B) qRT-PCR for miR-125a-5p relative expression in NSC-34 cells transfected with miRNA NC mimic or miR-125a-5p mimic. (C) Dual-luciferase reporter assays in NSC-34 cells with TCTN2 luciferase reporter construct (WT-TCTN2) or mutant TCTN2 reporter construct (MUT-TCTN2). (D) Relative expression of miR-125a-5p by qRT-PCR in NSC-34 cells exposed to 100 ng of LPS or control for 24 h. (E) qRT-PCR for miR-125a-5p expression in NSC-34 cells treated with LPS or LPS + TSIIA (8 μM). (F) TCTN2 expression by qRT-PCR in NSC-34 cells transfected with pc-NC or pc-TCTN2 (TCTN2 overexpression plasmid). (G) The level of miR-125a-5p by qRT-PCR in NSC-34 cells transfected with pc-NC, pc-TCTN2 (TCTN2 overexpression plasmid), pc-TCTN2+miRNA NC mimic or pc-TCTN2+miR-125a-5p mimic before LPS exposure. LPS: lipopolysaccharide, TSIIA: Tanshinone IIA. ∗ P < 0.05.

Article Snippet: The mouse motor neuron-like NSC-34 cell line was provided by Cedarlane (Burlington, NC, USA) and cultivated at 37 °C, 5% CO 2 in Dulbecco's modified Eagle Medium (Life Technologies, Scotland, UK) plus 1% penicillin/streptomycin (Life Technologies) and 10% fetal calf serum (Biosera, Boussens, France).

Techniques: Sequencing, Binding Assay, Mutagenesis, Quantitative RT-PCR, Expressing, Transfection, Luciferase, Construct, Over Expression, Plasmid Preparation

Journal: Regenerative Therapy

Article Title: Tanshinone IIA protects motor neuron-like NSC-34 cells against lipopolysaccharide-induced cell injury by the regulation of the lncRNA TCTN2/miR-125a-5p/DUSP1 axis

doi: 10.1016/j.reth.2023.03.007

Figure Lengend Snippet: Enforced expression of TCTN2 alleviates LPS-induced cell injury through miR-125a-5p. NSC-34 cells were transfected with pc-NC, pc-TCTN2 (TCTN2 overexpression plasmid), pc-TCTN2+miRNA NC mimic or pc-TCTN2+miR-125a-5p mimic and then exposed to 100 ng/mL of LPS for 24 h. (A) CCK-8 assay for cell viability. (B) Representative images showing a cell apoptosis assay and cell apoptosis by flow cytometry. (C) ELISA assay for TNF-α, IL-6 and IL-1β levels using the corresponding assay kit. (D) Representative images depicting a western blot assay and the levels of PCNA and Bcl-2 by western blot. (E) SOD activity using the assay kit. (F) MDA level using the assay kit. LPS: lipopolysaccharide. ∗ P < 0.05.

Article Snippet: The mouse motor neuron-like NSC-34 cell line was provided by Cedarlane (Burlington, NC, USA) and cultivated at 37 °C, 5% CO 2 in Dulbecco's modified Eagle Medium (Life Technologies, Scotland, UK) plus 1% penicillin/streptomycin (Life Technologies) and 10% fetal calf serum (Biosera, Boussens, France).

Techniques: Expressing, Transfection, Over Expression, Plasmid Preparation, CCK-8 Assay, Apoptosis Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Activity Assay

Journal: Regenerative Therapy

Article Title: Tanshinone IIA protects motor neuron-like NSC-34 cells against lipopolysaccharide-induced cell injury by the regulation of the lncRNA TCTN2/miR-125a-5p/DUSP1 axis

doi: 10.1016/j.reth.2023.03.007

Figure Lengend Snippet: TCTN2 targets miR-125a-5p to regulate the expression of DUSP1. (A) Sequence of miR-125a-5p, the binding sites for miR-125a-5p in DUSP1 3′UTR and the mutation of the target sites. (B) Dual-luciferase reporter assays in NSC-34 cells with DUSP1 3′UTR luciferase reporter construct (WT-DUSP1 3′UTR) or mutant DUSP1 3′UTR reporter construct (MUT-DUSP1 3′UTR). (C) Relative expression of DUSP1 protein by western blot in NSC-34 cells exposed to 100 ng/mL of LPS or control for 24 h. (D) Western blot for DUSP1 protein level in NSC-34 cells treated with LPS or LPS + TSIIA (8 μM). (E) DUSP1 protein level by western blot in NSC-34 cells transfected with si-NC or si-DUSP1. (F) qRT-PCR for miR-125a-5p expression in NSC-34 cells transfected with inhibitor NC or miR-125a-5p inhibitor. (G) Western blot for DUSP1 protein level in LPS-exposed NSC-34 cells transfected with inhibitor NC, miR-125a-5p inhibitor, miR-125a-5p inhibitor + si-NC or miR-125a-5p inhibitor + si-DUSP1. (H) DUSP1 protein level by western blot in LPS-treated NSC-34 cells transfected with pc-NC, pc-TCTN2 (TCTN2 overexpression plasmid), pc-TCTN2+miRNA NC mimic or pc-TCTN2+miR-125a-5p mimic. LPS: lipopolysaccharide, TSIIA: Tanshinone IIA. ∗ P < 0.05.

Article Snippet: The mouse motor neuron-like NSC-34 cell line was provided by Cedarlane (Burlington, NC, USA) and cultivated at 37 °C, 5% CO 2 in Dulbecco's modified Eagle Medium (Life Technologies, Scotland, UK) plus 1% penicillin/streptomycin (Life Technologies) and 10% fetal calf serum (Biosera, Boussens, France).

Techniques: Expressing, Sequencing, Binding Assay, Mutagenesis, Luciferase, Construct, Western Blot, Transfection, Quantitative RT-PCR, Over Expression, Plasmid Preparation

Journal: Regenerative Therapy

Article Title: Tanshinone IIA protects motor neuron-like NSC-34 cells against lipopolysaccharide-induced cell injury by the regulation of the lncRNA TCTN2/miR-125a-5p/DUSP1 axis

doi: 10.1016/j.reth.2023.03.007

Figure Lengend Snippet: The reduced expression of miR-125a-5p alleviates LPS-induced cell damage by up-regulating DUSP1. NSC-34 cells were transfected with inhibitor NC, miR-125a-5p inhibitor, miR-125a-5p inhibitor + si-NC or miR-125a-5p inhibitor + si-DUSP1 and then exposed to 100 ng/mL of LPS for 24 h. (A) Cell viability by CCK-8 assay. (B) Representative images showing a cell apoptosis assay and cell apoptosis by flow cytometry. (C) ELISA assay for TNF-α, IL-6 and IL-1β levels using the corresponding assay kit. (D) Representative images depicting a western blot assay and the levels of PCNA and Bcl-2 by western blot. (E) SOD activity using the assay kit. (F) MDA level using the assay kit. LPS: lipopolysaccharide. ∗ P < 0.05.

Article Snippet: The mouse motor neuron-like NSC-34 cell line was provided by Cedarlane (Burlington, NC, USA) and cultivated at 37 °C, 5% CO 2 in Dulbecco's modified Eagle Medium (Life Technologies, Scotland, UK) plus 1% penicillin/streptomycin (Life Technologies) and 10% fetal calf serum (Biosera, Boussens, France).

Techniques: Expressing, Transfection, CCK-8 Assay, Apoptosis Assay, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Western Blot, Activity Assay

Journal: Regenerative Therapy

Article Title: Tanshinone IIA protects motor neuron-like NSC-34 cells against lipopolysaccharide-induced cell injury by the regulation of the lncRNA TCTN2/miR-125a-5p/DUSP1 axis

doi: 10.1016/j.reth.2023.03.007

Figure Lengend Snippet: Tanshinone IIA protects LPS-induced NSC-34 cell injury by regulating the lncRNA TCTN2/miR-125a-5p/DUSP1 axis.

Article Snippet: The mouse motor neuron-like NSC-34 cell line was provided by Cedarlane (Burlington, NC, USA) and cultivated at 37 °C, 5% CO 2 in Dulbecco's modified Eagle Medium (Life Technologies, Scotland, UK) plus 1% penicillin/streptomycin (Life Technologies) and 10% fetal calf serum (Biosera, Boussens, France).

Techniques: