Journal:

Article Title: Cloning of the cDNA encoding the urotensin II precursor in frog and human reveals intense expression of the urotensin II gene in motoneurons of the spinal cord

doi:

Figure Lengend Snippet: Tissue distribution of human prepro-UII mRNA. (A) Dot blot analysis of prepro-UII mRNA expression in various human tissues. The master blot (CLONTECH) consisted of 50 human-tissue poly(A) RNAs (80–448 ng per dot) normalized by using the RNA expression levels of eight housekeeping genes. Positive controls consisted of human genomic DNA. Negative controls included yeast and Escherichia coli RNA or DNA, as well as human repetitive genomic sequences. The blot was probed with the human prepro-UII cDNA probe and exposed for 2 days onto an X-Omat film (B) Northern blot analysis of prepro-UII mRNA expression in the human spinal cord. Spinal cord poly(A) mRNA (2 μg) was hybridized with the human prepro-UII cDNA probe. The molecular weight was determined by using RNA markers. (C) X-ray autoradiographs showing the distribution of prepro-UII mRNA in the human spinal cord. Coronal sections were hybridized with the antisense (Top) or sense (Bottom) prepro-UII riboprobe and exposed for 10 days onto x-ray film.

Article Snippet: Thus, human spinal cord poly(A) RNA (CLONTECH) was used for amplification of the 5′-end of human UII cDNA with a commercially available rapid amplification of cDNA ends (RACE) kit (Marathon cDNA amplification kit, CLONTECH).

Techniques: Dot Blot, Expressing, RNA Expression, Genomic Sequencing, Northern Blot, Molecular Weight

Journal: PLoS ONE

Article Title: Development and characterization of sphingosine 1-phosphate receptor 1 monoclonal antibody suitable for cell imaging and biochemical studies of endogenous receptors

doi: 10.1371/journal.pone.0213203

Figure Lengend Snippet: S1P 1 is expressed in human spinal cord. Immunohistochemistry was performed in frozen human spinal cord sections using 2B9 and control IgG2A antibodies as described in methods section. Scale bar, 100 μm. A, S1P 1 is expressed in human kidney. Immunohistochemistry was performed in human TMA kidney samples using 2B9 and control IgG2A antibodies (1:50, w/v) as was previously described . Right panel shows 2mm TMA kidney sections, left panel shows X20 magnification.

Article Snippet: Frozen human spinal cord sections from BioChain Institute (Newark, USA) were post-fixed in 4% formaldehyde for 15 min at room temperature.

Techniques: Immunohistochemistry, Control

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