Journal: Nature neuroscience

Article Title: A glycolytic shift in Schwann cells supports injured axons

doi: 10.1038/s41593-020-0689-4

Figure Lengend Snippet: a-i, Representative immunofluorescence for the indicated metabolic components on longitudinal frozen sections from uninjured control nerve segments and axotomized distal sciatic nerve stumps at the shown post-injury times. HK2: Hexokinase 2 (a). GPI: Glucose-6-phosphate isomerase (b). ALDA: Aldolase A (c). GAPDH: Glyceraldehyde-3-phosphate dehydrogenase (d). PGK1: Phosphoglycerate kinase 1 (e). PGAM1: Phosphoglycerate mutase 1 (f). ENO1: Enolase 1 (g). PKM1: Pyruvate kinase M1 (h). LDHB: Lactate dehydrogenase B (i). Arrows depict colocalization in SCs. Scale bars: 50μm. The experiments for each component were reproduced three times independently with similar results.

Article Snippet: Following primary antibodies were used for nerve and teased fiber immunofluorescence: Neurofilament 200 (1:500, Sigma, N4142), Hexokinase I C35C4 (1:200, Cell Signaling, 2024), Hexokinase II C64G5 (1:200, Cell Signaling, 2867), GPI (1:200, Proteintech, 15171-1-AP), PFKM (1:200, Proteintech, 55028-1-AP), Aldolase A D73H4 (1:200, Cell Signaling, 8060), GAPDH (1:500, Sigma, G9545), PGK1 (1:150, Proteintech, 17811-1-AP), PGAM1 D3J9T (1:50, Cell Signaling, 12098), Enolase-1 D2S1A (1:200, Cell Signaling, 13410), PKM1 D30G6 (1:400, Cell Signaling, 7067), PKM2 D78A4 (1:200, Cell Signaling, 4053), PFKFB3 D7H4Q (1:200, Cell Signaling, 13123), LDHA/LDHC C28H7 (1:200, Cell Signaling, 3558), LDHB (1:200, Proteintech, 14824-1-AP), Glut1 D3J3A (1:200, Cell Signaling, 12939), PDHK1 C47H1 (1:100, Cell Signaling, 3820), Pyruvate Dehydrogenase C54G1 (1:100, Cell Signaling, 3205), CS (1:150, Proteintech, 16131-1-AP), IDH3A (1:100, Proteintech, 15909-1-AP), OGDH (1:50, Proteintech, 15212-1-AP), MCT1 M-45 (1:200, Santa Cruz Biotechnology, sc-50325), MCT4 H-90 (1:250, Santa Cruz Biotechnology, sc-50329), Phospho-S6 Ribosomal Protein (Ser240/244) D68F8 (1:800, Cell Signaling, 5364), Phospho-Akt (Ser473) D9E (1:100, Cell Signaling, 4060), HIF-1 alpha (1:200, Novus Biologicals, NB100-479), c-Myc D84C12 (1:800, Cell Signaling, 5605), c-Myc/N-Myc D3N8F (1:800, Cell Signaling, 13987), Phospho-AMPKα (Thr172) 40H9 (1:100, Cell Signaling, 2535), S100 4C4.9 (1:200, Thermo Fisher Scientific, MA5-12966), P0 (1:1000, Aves Labs, PZO), TUJ1/TUBB3 (1:500, BioLegend, 801202).

Techniques: Expressing, Immunofluorescence, Control

Journal: Nature neuroscience

Article Title: NG2 glia protect against prion neurotoxicity by inhibiting microglia-to-neuron prostaglandin E2 signaling.

doi: 10.1038/s41593-024-01663-x

Figure Lengend Snippet: Fig. 7 | PGE2 enhances prion neurotoxicity mainly through the EP4 receptor (Ptger4). a,b, Live-cell imaging (a) and quantitative analysis (b) of chronically prion-infected HovS cells expressing control (Ctrl) transgene or one of the four PGE2 receptors (Ptger1–4). Effects of PGE2 treatment on prion-induced cell toxicity were measured with the ratio of GFP signals under the PGE2 condition against the DMSO condition; n = 4 independent experiments. Data are presented as mean ± s.e.m. One-way ANOVA with Benjamini–Hochberg FDR adjustment for multiple comparisons: P < 0.0001 (Ptger1 versus Ctrl); P = 0.2246 (Ptger2 versus Ctrl); P = 0.3351 (Ptger3 versus Ctrl); P < 0.0001 (Ptger4 versus Ctrl). c, Immunofluorescence of NeuN, Map2 and Tau showing cellular damage of prion- infected primary neurons treated with different concentrations of Ptger4 agonist L902688. d, Quantification of neuronal density as well as Map2-positive and Tau positive areas shown in c; n = 6 independent experiments. Data are presented as

Article Snippet: 3 n atu re p o rtfo lio | rep o rtin g su m m ary A p ril 2 0 2 3 Materials & experimental systems n/a Involved in the study Antibodies Eukaryotic cell lines Palaeontology and archaeology Animals and other organisms Clinical data Dual use research of concern Plants Methods n/a Involved in the study ChIP-seq Flow cytometry MRI-based neuroimaging Antibodies Antibodies used mouse monoclonal antibody against actin (1:10,000, Merck Millipore, MAB1501R, clone C4); mouse monoclonal antibody against PrP (POM1, 1:5000, homemade); rabbit polyclonal antibody against NG2 (1:500, MERCK, AB5320); rabbit polyclonal antibody against PDGFRα (1:500, Santa Cruz, SC-338); rabbit monoclonal antibody against NeuN (1:1000, Abcam, ab177487, clone EPR12763); rabbit polyclonal antibody against NG2 (1:500, a gift from Prof. Stallcup); rabbit polyclonal antibody against Iba1 (1:500, Wako, 019-19741); Rat monoclonal antibody against Cd68 (1:200, BioRad, MCA1957, clone FA-11); rabbit polyclonal antibody against Map2 (1:200, Biolegend, 840601), mouse monoclonal antibody against Cox2 (1:200, Santa Cruz, sc-166475, clone D-12); mouse monoclonal antibody against Ptges (1:200, Santa Cruz, sc-365844, clone H-3); rabbit polyclonal antibody against EP1 (1:200, Bioss Antibodies, BS-6316R); rabbit monoclonal antibody against EP2 (1:200, Abcam, ab167171, clone EPR8030(B)); rabbit polyclonal antibody against EP3 (1:200, Cayman Chemical, 101760); mouse monoclonal antibody against EP4 (1:200, ProteinTech, 66921-1-Ig, clone 4A2A12); rat monoclonal antibody against CD11b antibody (30 ul in 10 ml, ThermoFisher Scientific, 14-0112-82, clone M1/70), mouse monoclonal antibody against Tau (1:200, ThermoFisher Scientific, MN1010, clone BT2), chicken polyclonal antibody against NeuN (1:1000, Merck, ABN91), goat polyclonal antibody against rat IgG (30 ul in 10 m, Jackson ImmunoResearch, 112-005-167); HRP-conjugated goat antirabbit IgG antibody (1:10,000, Jackson ImmunoResearch, 111-035-003); HRP-conjugated goat anti-mouse IgG antibody (1:10,000, Jackson ImmunoResearch, 115-035-003); Alexa488-conjugated goat anti-mouse IgG antibody (1:3000, ThermoFisher Scientific, A32723); Alexa594-conjugated goat anti-rabbit IgG antibody (1:3000, ThermoFisher Scientific, A32740); Alexa647-conjugated goat anti-chicken IgG antibody (1:3000, ThermoFisher Scientific, A32933).

Techniques: Live Cell Imaging, Infection, Expressing, Control, Immunofluorescence