Journal: The Journal of Neuroscience

Article Title: miR126-5p Downregulation Facilitates Axon Degeneration and NMJ Disruption via a Non–Cell-Autonomous Mechanism in ALS

doi: 10.1523/JNEUROSCI.3037-17.2018

Figure Lengend Snippet: miR126-5p is depleted in SOD1 G93A muscles and regulates Sema3 and NRP expression. A , NanoString chip screen heat map of significantly altered miRs in P60 muscles of SOD1 G93A compared with LM mice (extended table under ). Red and green represent a high or low abundance of miRs, respectively. * p < 0.05 (Student's t test; n = 3). B , miR126-5p was the most significantly downregulated miRNA in SOD1 G93A muscles (Student's t test; n = 3, ** p = 0.003). C , qPCR analysis of P60 GC muscle extracts further validates the decrease in miR 126-5p in SOD1 G93A ( n = 3). D–G , qPCR analysis of Sema3A, NRP1, Sema3B, and NRP2 transcript levels in HeLa cells overexpressing either miR126-5p or miR142 demonstrates a reduction in their expression levels specifically under miR126-5p overexpression (Student's t test; n = 3, * p = 0.0438, * p = 0.034, * p = 0.031, and * p = 0.0434, respectively). H , Representative TIRF images of U87MG cells reveal a detachment of the cell membrane from the culture dish surface after Sema3A is added to the culture medium. Scale bar, 10 μm. I , Impedance recording of live cells over time shows that U87MG cells overexpressing miR126-5p are unresponsive to Sema3A added to the culture medium because their impedance continuously increases, whereas the impedance of U87MG cells overexpressing miR142 decreases after treatment .

Article Snippet: Antibodies were used at the following concentrations: anti-Neurofilament Heavy Chain 1:500 (NFH, Abcam, ab72996; 1:1000), synaptophysin (Millipore, MAB5258; 1:300), synaptotagmin (Alomone Labs, ant-003; 1:300), anti-NRP1 (1:100), anti-Sema3A (1:100), anti-NRP2 (1:100), and anti-Sema3B (1:100).

Techniques: Expressing, Over Expression

Journal: Arthritis Research & Therapy

Article Title: Semaphorin 3G exacerbates joint inflammation through the accumulation and proliferation of macrophages in the synovium

doi: 10.1186/s13075-022-02817-7

Figure Lengend Snippet: Nrp2 expression in activated macrophages during joint inflammation. A Nrp2 expression in immune cells in the CIA synovium. The hind paws of CIA-induced mice were digested and subjected to flow-cytometric analysis. The representative histograms of Nrp2 staining and the cumulative data are shown. Data are expressed as the means ± SD. N = 3 from three independent experiments. B The characteristics of Nrp2-high macrophages. The representative histograms of CD86 and MHC class II expression on joint-infiltrating macrophages (left) and their cumulative data (right) are shown. N = 6 from two independent experiments. C Nrp2 expression on BMMs. BMMs were cultured under the indicated conditions, and Nrp2 expression was determined by flow cytometry. The representative histograms and the cumulative data are shown. N = 3 from three independent experiments. D NRP2 expression in synovium-infiltrating cells in RA. The deposited single-cell RNA-seq data were reanalyzed, and several cell types were defined by UMAP. NRP2 expression is denoted by purple dots. The statistical analyses were performed using an unpaired t-test

Article Snippet: Anti-neuropilin2 (Nrp2) polyclonal antibodies conjugated with FITC were purchased from Alomone Labs (#ANR-062).

Techniques: Expressing, Staining, Cell Culture, Flow Cytometry, RNA Sequencing