Journal: bioRxiv

Article Title: Atxn2 -CAG100-knock-in affects mouse lifespan and vestibulo-cerebellar function via neural disconnection

doi: 10.1101/333443

Figure Lengend Snippet: Dysregulations of protein and mRNA levels were observed for several factors involved in axon myelination, namely the axon-localized Neurofilament heavy / medium / light chain subunits (NEFH / NEFM / NEFL), the synapse-localized Neuroplastin (NPTN), the synapse-localized alpha-Synuclein (SNCA), versus the oligodendrocyte-localized Myelin Basic Protein (MBP), 2’,3’-Cyclic Nucleotide 3’ Phosphodiesterase (CNP), Proteolipid Protein (PLP1), Myelin oligodendrocyte glycoprotein (MOG) and Myelin associated glycoprotein (MAG). The abundance of the protein of interest was normalized versus beta-Actin (ACTB) as total tissue loading control, and separately versus Calbindin-1 (CALB1) as marker of Purkinje neuron somatodendritic compartment preservation. The mRNA abundance of each factor is shown with normalization versus TATA-box-binding protein ( Tbp ) mRNA, to elucidate if the dysregulation might be triggered at the transcriptional level by the RNA-binding ATXN2 expansion via direct interaction, or represents a post-transcriptional event. T-test was used in general.

Article Snippet: The TaqMan ® Assays utilized for this study are: Atxn2 (Mm01199894_m1), Calb1 (Mm00486647-m1), Cnp (Mm01306641_m1), Hapln1 (Mm00618325_m1), Hapln2 (Mm00480745_m1), Hapln3 (Mm00724203_m1), Hapln4 (Mm00625974_m1), Hprt1 (Mm00446968_m1), Klk6 (Mm00478322_m1), Mag (Mm00487538_m1), Mal (Mm01339780_m1), Mbp (Mm01266402_m1), Mobp long transcripts (Mm02745649_m1), Mobp short transcripts (Mm01348317_g1), Mog (Mm00447824_m 1), Nefh (Mm01191456_m1), Nefl (Mm01315666_m1), Nefm (Mm00456201_m1), Nptn (Mm00485990_m1), Plp1 (Mm01297210_m1), Prnp (Mm004483 89_m 1), Rgs8 (Mm01290239_m1), Rtn4 (Mm00445861_m1), Snca (Mm00447333_m1), Tbp (Mm00446973_m1), Tuba4a (Mm00849767_s1).

Techniques: Marker, Preserving, Binding Assay, RNA Binding Assay

Journal: Frontiers in Immunology

Article Title: High serum prevalence of autoreactive IgG antibodies against peripheral nerve structures in patients with neurological post-COVID-19 vaccination syndrome

doi: 10.3389/fimmu.2024.1404800

Figure Lengend Snippet: Antibody target identification and confirmation in PCVS sera. Volcano plot representing significantly enriched proteins (labeled in red) in patient #12 IgG IP (A) and patient #41 IgG IP (B) compared to a negative control; in A-B: the x-axis displays the log2-transformed fold change, and the y-axis represents the -log10-transformed p value. (C) Cell-based assays with patient #12 serum testing IgG reactivity against neurofilament subunits and control HEK293 cells. (D) Costaining of sciatic nerve teased fibers with a commercial NF-H antibody (D’) and patient #12 serum (D’’) showing clear signal overlap (D’’’). (E) ELISA analysis of DFS-70 and patient #41 serum. PC: positive control serum. NC: negative control serum. The standard reference serum OD was 0.278 (dotted line). OD: optical density. (F) Hep2 staining of patient #41 serum resembling fine speckled nuclear staining typical of DFS-70 IgGs.

Article Snippet: In brief, commercially available plasmids encoding Neurofilament heavy chain (NFH; #RC213487 Origene, Rockville, MD, USA), Neurofilament medium chain (NFM; #RC224475 Origene), Neurofilament light chain (NFL; #RC205920 Origene), Peripherin (PRPH; #RC207561 Origene), and Alpha-Internexin (INA; #RC202877 Origene) were transiently expressed in HEK293 cells using PEI-mediated transfection on polylysine-coated coverslips.

Techniques: Drug discovery, Labeling, Negative Control, Transformation Assay, Control, Enzyme-linked Immunosorbent Assay, Positive Control, Staining

Journal: Biomedicines

Article Title: Comparison of the Confluence-Initiated Neurogenic Differentiation Tendency of Adipose-Derived and Bone Marrow-Derived Mesenchymal Stem Cells

doi: 10.3390/biomedicines9111503

Figure Lengend Snippet: Neuron-related gene expressions of the ADSCs during confluence-initiated differentiation. The time course experiments of detecting neuron-associated gene expressions include class III β-tubulin ( TUBB3 ) ( A ), microtubule-associated protein 2 ( MAP2 ) ( B ), neurofilament medium polypeptide ( NEFM ) ( C ), and neurofilament heavy polypeptide ( NEFH ) ( D ) in the spontaneously differentiated ADSCs after high confluence was achieved in the cultures by RT-qPCR. The value of each gene expression was normalized to the expression of glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ). Relative fold changes of the values from day 1 to day 7 were compared with the value of day 0 (without induction) as one. The values are expressed as mean ± S.D. with three experimental replicates. Statistical significance was determined using the Mann–Whitney U test. “*” represents p < 0.05.

Article Snippet: Primary antibodies against microtubule-associated protein 2 (MAP2; GeneTex Inc., Irvine, CA, USA), class III β-tubulin (TUBB3; Santa Cruz Biotechnology, Santa Cruz, CA, USA), neurofilament medium polypeptide (NEFM; OriGene Technologies, Medical Center Drive, Rockville, MD, USA), and neurofilament light polypeptide (NEFL; OriGene Technologies) were added on the slides at an appropriate dilution for IF staining.

Techniques: Quantitative RT-PCR, Gene Expression, Expressing, MANN-WHITNEY

Journal: Biomedicines

Article Title: Comparison of the Confluence-Initiated Neurogenic Differentiation Tendency of Adipose-Derived and Bone Marrow-Derived Mesenchymal Stem Cells

doi: 10.3390/biomedicines9111503

Figure Lengend Snippet: Comparison of neuron-related gene expressions of the ADSCs and BMSCs during confluence-initiated neurogenic differentiation. The time course experiments of detecting neuron-associated gene expressions include class III β-tubulin ( TUBB3 ) ( A ), microtubule-associated protein 2 ( MAP2 ) ( B ), neurofilament medium polypeptide ( NEFM ) ( C ), and neurofilament heavy polypeptide ( NEFH ) ( D ) in the spontaneously differentiated ADSCs after high confluence was achieved in the cultures by RT-qPCR. The value of each gene expression was normalized to the expression of glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ). Relative fold changes of the values from day 1 to day 7 were compared with the value of day 0 (without induction) as one. The values are expressed as mean ± S.D. with three experimental replicates. Statistical significance to compare confluence-initiated differentiation of the ADSCs and BMSCs was determined using one-way ANOVA analysis. “*” represents p < 0.05. “**” represents p < 0.01. “***” represents p < 0.001.

Article Snippet: Primary antibodies against microtubule-associated protein 2 (MAP2; GeneTex Inc., Irvine, CA, USA), class III β-tubulin (TUBB3; Santa Cruz Biotechnology, Santa Cruz, CA, USA), neurofilament medium polypeptide (NEFM; OriGene Technologies, Medical Center Drive, Rockville, MD, USA), and neurofilament light polypeptide (NEFL; OriGene Technologies) were added on the slides at an appropriate dilution for IF staining.

Techniques: Comparison, Quantitative RT-PCR, Gene Expression, Expressing

Journal: Biomedicines

Article Title: Comparison of the Confluence-Initiated Neurogenic Differentiation Tendency of Adipose-Derived and Bone Marrow-Derived Mesenchymal Stem Cells

doi: 10.3390/biomedicines9111503

Figure Lengend Snippet: Comparison of neuron-related protein expressions of the ADSCs and BMSCs during confluence-initiated neurogenic differentiation. ( A , E , I ) Class III β-tubulin (TUBB3), ( B , F , J ) microtubule-associated protein 2 (MAP2), ( C , G , K ) neurofilament medium polypeptide (NEFM), and ( D , H , L ) neurofilament light polypeptide (NEFL) of the factor-induced ADSCs, confluence-initiated ADSCs, and confluence-initiated BMSC differentiated neuron-like cells on day 7 detected by immunofluorescence staining are shown in green fluorescence. DNA stained by DAPI is shown in blue fluorescence (magnification × 100; the scale bar = 100 µm). ( M ) The mean of the total immunofluorescence intensity from a fixed number of cells in six fields of one slide was quantified by the Image-Pro Plus v4.5.0.29. ( N ) The mean of the value that was normalized to the cell numbers in the field was calculated. All the values are expressed as mean ± S.D. with three experimental replicates. Statistical significance to compare the multiple groups was determined using one-way ANOVA analysis. “*” represents p < 0.05. “**” represents p < 0.01.

Article Snippet: Primary antibodies against microtubule-associated protein 2 (MAP2; GeneTex Inc., Irvine, CA, USA), class III β-tubulin (TUBB3; Santa Cruz Biotechnology, Santa Cruz, CA, USA), neurofilament medium polypeptide (NEFM; OriGene Technologies, Medical Center Drive, Rockville, MD, USA), and neurofilament light polypeptide (NEFL; OriGene Technologies) were added on the slides at an appropriate dilution for IF staining.

Techniques: Comparison, Immunofluorescence, Staining, Fluorescence

Journal: International journal of andrology

Article Title: Neurotrophic effect of bone marrow mesenchymal stem cells for erectile dysfunction in diabetic rats.

doi: 10.1111/j.1365-2605.2012.01250.x

Figure Lengend Snippet: Figure 4 Neurofilament content in dorsal nerve. Representative images of each experimental group are shown in above panel. Original magnifi- cation is 400·. Result of neurofilament (NF) quantification is expressed as the content of NF ⁄ area of the nerve (in pixel). C: normal control group, DM: diabetic group, MSC: diabetic rats injected with bone marrow-derived mesenchymal stem cells (BM-MSCs), Medium: diabetic rats injected with BM-MSCs-conditioned medium. *p < 0.05 compared with DM group.

Article Snippet: The primary antibodies used in the present study were: rabbit anti neuronal nitric oxide synthase (nNOS; Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA, 1 : 200) and rabbit anti neurofilament (NF; Boster, Wuhan, Hubei, China, 1 : 200).

Techniques: Control, Injection, Derivative Assay