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 membranes were blocked in 5% BSA/TBS-T, and incubated overnight at 4 °C with primary antibodies against 1C2 (Chemicon #MAB1574), ATXN2 (monoclonal from BD Biosciences #611378, 1:500; polyclonal from Proteintech #21776-1-AP, 1:500), ACTB (Sigma #A5441, 1:10000), CALB1 (Cell Signaling #13176, 1:2500), CNP (Cell Signaling #5664S, 1:1000), MAG (Cell Signaling #9043S, 1:500), MBP (Merck #05-675, 1:250), MOG (Abcam #ab32760, 1:500), NEFH/M (Proteintech #18934-1-AP, 1:500), NEFL (Cell Signaling #2837S, 1:1000), NPTN (Alomone Labs #ANR-090, 1:500), PLP1 (Abcam #ab28486, 1:1000), SNCA (Cell Signaling #2642S, 1:1000), TUBA4A (Aviva Systems #ARP40179-P050, 1:500).

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

Journal: Neuroscience Bulletin

Article Title: Rapid and Sparse Labeling of Neurons Based on the Mutant Virus-Like Particle of Semliki Forest Virus

doi: 10.1007/s12264-019-00362-z

Figure Lengend Snippet: Mutant SFV VLP rapidly and sparsely labels Purkinje neurons in vivo. To sparsely label Purkinje cells, the mutant virus was injected into the third cerebellar lobule (3Cb) region. The mice were sacrificed at 24 hpi and sagittal and coronal sections were prepared. A The EGFP signals were co-localized with the Purkinje cell marker calbindin-D28k (arrows). B The fine structure of the Purkinje cells was apparent, including the dendritic tree, long axon, and axonal branches (asterisks). These images are representative of 15 sections (n = 3 mice).

Article Snippet: To stain Purkinje cells, fixed sagittal sections 80 μm thick were immunostained with a mouse monoclonal antibody against calbindin-D28k (Proteintech) and amplified with a Cy3-conjugated anti-mouse secondary antibody (Merck).

Techniques: Mutagenesis, In Vivo, Virus, Injection, Marker

Journal: PLOS Pathogens

Article Title: Deciphering infected cell types, hub gene networks and cell-cell communication in infectious bronchitis virus via single-cell RNA sequencing

doi: 10.1371/journal.ppat.1012232

Figure Lengend Snippet: (a) White arrows indicate the localization of IBV N protein in AQP2-expressing collecting duct cells. Green fluorescence shows positive staining for IBV N and red fluorescence shows staining for AQP2. (b) White arrows indicate the localization of IBV N protein in CALB1-expressing distal tubule cells. Red fluorescence shows positive staining for IBV N and green fluorescence shows staining for CALB1. (c) Changes in cell communication numbers: The top network diagram shows cell clusters as nodes, with line thickness indicating changes in communication numbers. The lower heatmap details these changes, with rows representing signal-sending cells and columns indicating signal-receiving cells. The color scale reflects the inter-group differences in signal communication frequency between different cell types (number of communications in the infected group—number in control group). The bar plots at the top and right side represent the overall differences in the number of signals sent/received by specific cell clusters. (d) Changes in cell communication strength: Similar to (c), with the top network diagram displaying changes in communication strength (communication strength in the infected group—strength in the control group). In the lower heatmap, the color scale reflects the inter-group differences in signal communication strength between different cell types. The bar plots at the top and right side represent the overall differences in the strength of signals sent/received by specific cell clusters (infected group—control group). (e) Inter-group differences in the communication strength of specific signaling pathways (receptor-ligand pairs) across cell clusters. Rows represent signal pathways and columns correspond to cell clusters, with heatmap colors depicting the strength variation of signals (infect group vs. control group). Upper left triangles for signal sent and lower right triangles for signal received. The bar plot on the right side shows the overall difference in communication strength of these signaling pathways between the IBV-infected group and the control group.

Article Snippet: mouse mab calb1 , 1:200 , Boster Bio (BM0203).

Techniques: Expressing, Fluorescence, Staining, Infection, Control, Protein-Protein interactions

Journal: PLOS Pathogens

Article Title: Deciphering infected cell types, hub gene networks and cell-cell communication in infectious bronchitis virus via single-cell RNA sequencing

doi: 10.1371/journal.ppat.1012232

Figure Lengend Snippet: Antbodies and reagents used in present study.

Article Snippet: mouse mab calb1 , 1:200 , Boster Bio (BM0203).

Techniques: Blocking Assay, Immunofluorescence