human spinal cord samples  (Radboud University)

Journal: Journal of molecular neuroscience : MN

Article Title: Alpha-Synuclein Loss in Spinal Muscular Atrophy

doi: 10.1007/s12031-010-9422-1

Figure Lengend Snippet: qRT-PCR of SMN and SNCA in human SMA fibroblasts and NSC-34 cell clones. a qRT-PCR of SMN and SNCA in human SMA fibroblasts. Fibroblasts derived from an SMA type I and type III patient were compared to control. SMN1 (left) showed a decreased expression from control to type III to type I, respectively. SNCA mRNA level (right) showed a similar deceasing profile across samples from the same patients. Regression analysis using the RT-PCR cycle counts indicated a significant proportion of the variance in SNCA can be explained by knowing the SMN1 level (R2 = 0.49 (F(1,7) = 8.8049, p < 0.02). b qRT-PCR of Smn and Snca in NSC-34 cell clones. NSC-34 cell clones with differing SMN expression level, including that used in Table 1, were examined at passage 7. The C-6 cells Smn expression was 20% lower than control, while Snca expression was 48% lower. Smn expression in the more severe C-2 cells was lowered by 85% compared to control, while Snca expression was 98% lower. All measurements in this figure represent averaged triplicates

Article Snippet: Human SMA Fibroblast and Spinal Cord Samples Human untransformed fibroblasts of an SMA type 1 patient, an SMA carrier (SMA patient's mother), and a healthy control were obtained from Coriell Institute tissue repository (Camden, NJ, USA).

Techniques: Quantitative RT-PCR, Clone Assay, Derivative Assay, Control, Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: Journal of molecular neuroscience : MN

Article Title: Alpha-Synuclein Loss in Spinal Muscular Atrophy

doi: 10.1007/s12031-010-9422-1

Figure Lengend Snippet: SNCA protein analysis in cells with low Smn levels. Proteins samples from stable and transient transfected NSC-34 cells and patient fibroblasts were incubated with polyclonal rabbit anti-SNCA and anti-β-actin antibodies. The intensities were measured, and SNCA/β-actin ratios were compared. Intensities for the weaker monomer and stronger dimer band were quantified from the same stable clones used in Fig. 2; NSC-34 cells transfected with a control vector or shSMN plasmid 72 h previously; and patient fibroblasts from controls, an SMA type III patient, the carrier mother of an SMA type I patient, and the patient. This figure is representative of more than three blots that were run for these data

Article Snippet: Human SMA Fibroblast and Spinal Cord Samples Human untransformed fibroblasts of an SMA type 1 patient, an SMA carrier (SMA patient's mother), and a healthy control were obtained from Coriell Institute tissue repository (Camden, NJ, USA).

Techniques: Transfection, Incubation, Clone Assay, Control, Plasmid Preparation

Journal: Journal of molecular neuroscience : MN

Article Title: Alpha-Synuclein Loss in Spinal Muscular Atrophy

doi: 10.1007/s12031-010-9422-1

Figure Lengend Snippet: Synaptic vesicular genes in spinal cord of SMA type I patients and NSC-34 cell clones. a qRT-PCR of SV2A and SYN2 in human SMA fibroblasts. Fibroblasts derived from a control and two SMA type I patients were compared. Both vesicular transport genes showed lower expression in both type I patients. b qRT-PCR of Sv2a and Syn2 in NSC-34 clones. NSC-34 clones with differing SMN level were examined at passage 7. C-6 and C-2 cells data showed differentially decreased expression of Sv2a and Syn2 expression relative to controls. All represent averaged triplicates

Article Snippet: Human SMA Fibroblast and Spinal Cord Samples Human untransformed fibroblasts of an SMA type 1 patient, an SMA carrier (SMA patient's mother), and a healthy control were obtained from Coriell Institute tissue repository (Camden, NJ, USA).

Techniques: Clone Assay, Quantitative RT-PCR, Derivative Assay, Control, Expressing