nsc34 neurons  (Cedarlane)

Journal: Bioactive Materials

Article Title: Development of a PTEN -siRNA activated scaffold to promote axonal regrowth following spinal cord injury

doi: 10.1016/j.bioactmat.2026.01.022

Figure Lengend Snippet: PTEN -siRNA nanoparticle transfection enables efficient gene silencing with minimal impact on neuron health (A) PTEN -siRNA nanoparticles (100 nM) demonstrated highly efficient knockdown of target PTEN mRNA in differentiated NSC34 neurons, with peak knockdown on Day 3 for the delivered siRNA [t(2) = −4.840, p = 0.02007; one-tailed] with expression levels returning to baseline by Day 7 [t(2) = 0.05373, p = 0.5190; one-tailed]. RNA fold change and associated error was assessed using the 2 −ΔΔCt method and normalized to GAPDH as per (B) Treatment of neurons with PTEN- siRNA nanoparticles at 100 nM had a non-significant effect on the metabolic activity at 3 [t(2) = 0.6370, p = 0.5893, 95 % CI 74.4820 0.1; • = p ≤ 0.1, ∗ = p ≤ 0.05, ∗∗ = p ≤ 0.01, ∗∗∗ = p ≤ 0.001, ∗∗∗∗ = p ≤ 0.0001.

Article Snippet: NSC34 neuronal culture and transfections: NSC34 neurons (Cedarlane Cat No. CLU140), a hybrid cell line produced by the fusion of motor neuron enriched embryonic mouse spinal cord cells with mouse neuroblastoma cells, were cultured in growth medium Dulbecco's Modified Eagle Medium (DMEM; Sigma-Aldrich, Ireland) supplemented with 10 % fetal bovine serum (FBS; Labtech UK), 1 % (v/v) L-Glutamine (Sigma-Aldrich, Ireland), and 1 % (v/v) Penicillin- Streptomycin Solution (Sigma-Aldrich, Ireland) in a T-175 cell culture flask (37 °C, 5 % CO 2 ).

Techniques: Transfection, Knockdown, One-tailed Test, Expressing, Activity Assay, Control, Staining

Journal: Bioactive Materials

Article Title: Development of a PTEN -siRNA activated scaffold to promote axonal regrowth following spinal cord injury

doi: 10.1016/j.bioactmat.2026.01.022

Figure Lengend Snippet: Scaffold-mediated siRNA delivery does not affect the viability of neurons, results in prolonged downregulation of PTEN, and stimulates increased expression of BCL2 and GAP43 mRNA . (A) The metabolic activity of NSC34 neurons cultured on PTEN -siRNA activated scaffolds peaked by day 3 post-transfection [t(2) = 4.0599, p = 0.05565, 95 % CI 93.954 0.1; • = p ≤ 0.1, ∗ = p ≤ 0.05, ∗∗ = p ≤ 0.01, ∗∗∗ = p ≤ 0.001, ∗∗∗∗ = p ≤ 0.0001.

Article Snippet: NSC34 neuronal culture and transfections: NSC34 neurons (Cedarlane Cat No. CLU140), a hybrid cell line produced by the fusion of motor neuron enriched embryonic mouse spinal cord cells with mouse neuroblastoma cells, were cultured in growth medium Dulbecco's Modified Eagle Medium (DMEM; Sigma-Aldrich, Ireland) supplemented with 10 % fetal bovine serum (FBS; Labtech UK), 1 % (v/v) L-Glutamine (Sigma-Aldrich, Ireland), and 1 % (v/v) Penicillin- Streptomycin Solution (Sigma-Aldrich, Ireland) in a T-175 cell culture flask (37 °C, 5 % CO 2 ).

Techniques: Expressing, Activity Assay, Cell Culture, Transfection, Control, Lactate Dehydrogenase Assay, Gene Expression, One-tailed Test

Journal: Brain research bulletin

Article Title: RBM5 induces motor neuron apoptosis in hSOD1 G93A -related amyotrophic lateral sclerosis by inhibiting Rac1/AKT pathways.

doi: 10.1016/j.brainresbull.2024.111049

Figure Lengend Snippet: Fig. 3. Overexpression of RBM5 induces motor neurons apoptosis. (A) Western blotting analysis and qRT-PCR were performed to assess the efficiency of RBM5 overexpression (n=3). (B) Flow cytometry was employed to assess the apoptosis of NSC34 cells following transfection with RBM5 or EV plasmid for 48 h (n=3). (C) TUNEL staining of NSC34 cells after treatment with RBM5 or EV plasmid (n=3). (D) Cell viability was quantified using CCK-8 (n=5). Data are presented as means ± SD. Statistical analysis was performed using student’s t-test. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: The NSC34 cell line (Cedarlane Laboratories, Vancouver, Canada) is a hybrid of embryonic mouse neuroblastoma and spinal cord motor neurons.

Techniques: Over Expression, Western Blot, Quantitative RT-PCR, Flow Cytometry, Transfection, Plasmid Preparation, TUNEL Assay, Staining, CCK-8 Assay

Journal: International Journal of Molecular Sciences

Article Title: Specific Post-Translational Modifications of VDAC3 in ALS-SOD1 Model Cells Identified by High-Resolution Mass Spectrometry

doi: 10.3390/ijms232415853

Figure Lengend Snippet: Sequence coverage map of VDAC3 from NSC34, NSC34-SOD1WT and NSC34-SOD1G93A cell lines obtained by tryptic and chymotryptic digestion. The solid line indicates the sequence that was obtained from tryptic peptides; dotted lines: sequence obtained from chymotryptic peptides. Unique tryptic (indicated in bold and blue) and chymotryptic (indicated in bold and black) peptides originating from missed cleavages were used to distinguish and cover the sequences shared by isoforms. Sequences shared by multiple isoforms are shown in red. Sequence numbering considered the starting methionine residue, which is eliminated during protein maturation.

Article Snippet: NSC34 cell lines (mouse motor neuron-like) obtained from CELLutions Biosystem Inc. were stably transfected with pTet-ON plasmid (Clontech, Mountain View, CA, USA) harboring sequences encoding human SOD1 wt (NSC34-SOD1WT) or G93A (NSC34-SOD1G93A) and used as non-ALS motor neuron line and ALS motor neuron line, respectively [ ].

Techniques: Sequencing, Residue

Journal: International Journal of Molecular Sciences

Article Title: Specific Post-Translational Modifications of VDAC3 in ALS-SOD1 Model Cells Identified by High-Resolution Mass Spectrometry

doi: 10.3390/ijms232415853

Figure Lengend Snippet: Ox/Red ratio (average and standard deviation) of the absolute intensities of the molecular ions of sulfur containing tryptic peptides found in the analysis of VDAC3 from NSC34, NSC34-SOD1WT, NSC34-SOD1G93A cell lines reduced with DTT, carboxyamidomethylated and digested in-solution.

Article Snippet: NSC34 cell lines (mouse motor neuron-like) obtained from CELLutions Biosystem Inc. were stably transfected with pTet-ON plasmid (Clontech, Mountain View, CA, USA) harboring sequences encoding human SOD1 wt (NSC34-SOD1WT) or G93A (NSC34-SOD1G93A) and used as non-ALS motor neuron line and ALS motor neuron line, respectively [ ].

Techniques: Standard Deviation, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Specific Post-Translational Modifications of VDAC3 in ALS-SOD1 Model Cells Identified by High-Resolution Mass Spectrometry

doi: 10.3390/ijms232415853

Figure Lengend Snippet: Ox/Red ratio (average and standard deviation) of the absolute intensities of the molecular ions of sulfur containing chymotryptic peptides found in the analysis of VDAC3 from NSC34, NSC34-SOD1WT, NSC34-SOD1G93A cell lines reduced with DTT, carboxyamidomethylated and digested in-solution.

Article Snippet: NSC34 cell lines (mouse motor neuron-like) obtained from CELLutions Biosystem Inc. were stably transfected with pTet-ON plasmid (Clontech, Mountain View, CA, USA) harboring sequences encoding human SOD1 wt (NSC34-SOD1WT) or G93A (NSC34-SOD1G93A) and used as non-ALS motor neuron line and ALS motor neuron line, respectively [ ].

Techniques: Standard Deviation, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Specific Post-Translational Modifications of VDAC3 in ALS-SOD1 Model Cells Identified by High-Resolution Mass Spectrometry

doi: 10.3390/ijms232415853

Figure Lengend Snippet: MS/MS spectrum of the doubly charged molecular ion at m/z 660.2664 (calculated 660.2663) of the N-terminal tryptic peptide of VDAC3 from NSC34-SOD1WT cell line with cysteine residue 2 in the form of sulfonic acid and cysteine residue 8 in the carboxyamidomethylated form. The inset shows the full scan mass spectrum of the molecular ion. Fragment ions originating from the neutral loss of H 2 O are indicated by an asterisk. The fragment ion originating from the neutral loss of NH 3 is indicated by two asterisks.

Article Snippet: NSC34 cell lines (mouse motor neuron-like) obtained from CELLutions Biosystem Inc. were stably transfected with pTet-ON plasmid (Clontech, Mountain View, CA, USA) harboring sequences encoding human SOD1 wt (NSC34-SOD1WT) or G93A (NSC34-SOD1G93A) and used as non-ALS motor neuron line and ALS motor neuron line, respectively [ ].

Techniques: Tandem Mass Spectroscopy, Residue

Journal: International Journal of Molecular Sciences

Article Title: Specific Post-Translational Modifications of VDAC3 in ALS-SOD1 Model Cells Identified by High-Resolution Mass Spectrometry

doi: 10.3390/ijms232415853

Figure Lengend Snippet: Ox/Red ratio (average and standard deviation) of the absolute intensities of the molecular ions of tryptic peptides containing succinated and non-succinated cysteines found in the analysis of VDAC3 from NSC34, NSC34-SOD1WT, NSC34-SOD1G93A cell lines reduced with DTT, carboxyamidomethylated and digested in-solution.

Article Snippet: NSC34 cell lines (mouse motor neuron-like) obtained from CELLutions Biosystem Inc. were stably transfected with pTet-ON plasmid (Clontech, Mountain View, CA, USA) harboring sequences encoding human SOD1 wt (NSC34-SOD1WT) or G93A (NSC34-SOD1G93A) and used as non-ALS motor neuron line and ALS motor neuron line, respectively [ ].

Techniques: Standard Deviation, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Specific Post-Translational Modifications of VDAC3 in ALS-SOD1 Model Cells Identified by High-Resolution Mass Spectrometry

doi: 10.3390/ijms232415853

Figure Lengend Snippet: Ox/Red ratio (average and standard deviation) of the absolute intensities of the molecular ions of tryptic peptides containing deamidated and non-deamidated asparagine found in the analysis of VDAC3 from NSC34-SOD1G93A cell line reduced with DTT, carboxyamidomethylated and digested in-solution.

Article Snippet: NSC34 cell lines (mouse motor neuron-like) obtained from CELLutions Biosystem Inc. were stably transfected with pTet-ON plasmid (Clontech, Mountain View, CA, USA) harboring sequences encoding human SOD1 wt (NSC34-SOD1WT) or G93A (NSC34-SOD1G93A) and used as non-ALS motor neuron line and ALS motor neuron line, respectively [ ].

Techniques: Standard Deviation, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Specific Post-Translational Modifications of VDAC3 in ALS-SOD1 Model Cells Identified by High-Resolution Mass Spectrometry

doi: 10.3390/ijms232415853

Figure Lengend Snippet: MS/MS spectrum of the doubly charged molecular ion at m/z 924.9586 (calculated 924.9582) of the VDAC3 tryptic peptide from NSC34-SOD1G93A cell line containing the asparagine residue 215 in the deamidated form. The inset shows the full scan mass spectrum of molecular ion. Fragment ions originated from the neutral loss of H 2 O are indicated by an asterisk. Fragment ions originated from the neutral loss of NH 3 are indicated by two asterisks.

Article Snippet: NSC34 cell lines (mouse motor neuron-like) obtained from CELLutions Biosystem Inc. were stably transfected with pTet-ON plasmid (Clontech, Mountain View, CA, USA) harboring sequences encoding human SOD1 wt (NSC34-SOD1WT) or G93A (NSC34-SOD1G93A) and used as non-ALS motor neuron line and ALS motor neuron line, respectively [ ].

Techniques: Tandem Mass Spectroscopy, Residue

Journal: International Journal of Molecular Sciences

Article Title: Specific Post-Translational Modifications of VDAC3 in ALS-SOD1 Model Cells Identified by High-Resolution Mass Spectrometry

doi: 10.3390/ijms232415853

Figure Lengend Snippet: Ox/Red ratio (average and standard deviation) of the absolute intensities of the molecular ions of tryptic peptides containing succinimide intermediate found in the analysis of VDAC3 from NSC34, NSC34-SOD1WT, NSC34-SOD1G93A cell lines reduced with DTT, carboxyamidomethylated and digested in-solution.

Article Snippet: NSC34 cell lines (mouse motor neuron-like) obtained from CELLutions Biosystem Inc. were stably transfected with pTet-ON plasmid (Clontech, Mountain View, CA, USA) harboring sequences encoding human SOD1 wt (NSC34-SOD1WT) or G93A (NSC34-SOD1G93A) and used as non-ALS motor neuron line and ALS motor neuron line, respectively [ ].

Techniques: Standard Deviation, Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Specific Post-Translational Modifications of VDAC3 in ALS-SOD1 Model Cells Identified by High-Resolution Mass Spectrometry

doi: 10.3390/ijms232415853

Figure Lengend Snippet: Retention time, experimentally measured and calculated monoisotopic m/z of the molecular ions, position in the sequence, absolute intensities and peptide sequence of tryptic fragment containing asparagine residue 76 in succinimide intermediate form and asparagine residue 79 in isoaspartate methyl ester form found in the analysis of VDAC3 from NSC34 and NSC34-SOD1WT cell lines reduced with DTT, carboxyamidomethylated and digested in-solution.

Article Snippet: NSC34 cell lines (mouse motor neuron-like) obtained from CELLutions Biosystem Inc. were stably transfected with pTet-ON plasmid (Clontech, Mountain View, CA, USA) harboring sequences encoding human SOD1 wt (NSC34-SOD1WT) or G93A (NSC34-SOD1G93A) and used as non-ALS motor neuron line and ALS motor neuron line, respectively [ ].

Techniques: Sequencing, Residue