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. 1 | NG2 glia activation in prion disease models. a,b, Western blots (a) and quantification (b) of NG2 and NeuN in Tga20 COCS exposed to prions or NBH; n = 6 samples per condition. Data are presented as mean ± s.e.m. Unpaired t-test (two-sided): P < 0.0001 for NeuN; P = 0.0021 for NG2. c,d, Western blots (c) and quantification (d) of NG2 and Pdgfrα in brain tissues of mice inoculated with prions or NBH; n = 6 mice per condition. Data are presented as mean ± s.e.m. Unpaired t-test (two-sided): P = 0.0202 for Pdgfrα; P < 0.0001 for NG2. e, NG2 immunofluorescence showing NG2 glia activation in prion-infected Tga20 COCS

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: Activation Assay, Western Blot, Immunofluorescence, Infection

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. 3 | Loss of NG2 glia upregulates Cox2–Ptges expression. a,b, NG2 immunofluorescence (a) and quantification (b) in intact (DMSO) and NG2-glia- depleted (CP673451) C57BL/6J COCS; n = 7 slices per condition. Data are presented as mean ± s.e.m. Unpaired t-test (two-sided): P < 0.0001. c, qRT-PCR results showing downregulation of NG2 and Pdgfrα and upregulation of Cox2 and Ptges in NG2-glia-depleted (CP673451) C57BL/6J COCS; n = 6 samples. Data are presented as mean ± s.e.m. Multiple unpaired t-tests with Benjamini–Hochberg FDR adjustment for multiple comparisons. NG2: P < 0.0001; Pdgfrα: P < 0.0001; Cox2: P < 0.0001; Ptges: P = 0.0067. d,e, NG2 immunofluorescence (d) and quantification (e) in control (Ctrl) and PDGFAA-treated C57BL/6J COCS; n = 7 slices per condition. Data are presented as mean ± s.e.m. Unpaired t-test (two-sided): P < 0.0001. f, qRT-PCR results showing upregulation of NG2 and Pdgfrα and downregulation of Cox2 and Ptges in C57BL/6J COCS with increased NG2 glia density (PDGFAA) compared with NG2-glia-intact (Ctrl) C57BL/6J COCS; n = 4 samples for Ctrl; n = 6 samples for PDGFAA. Data are presented as mean ± s.e.m. Multiple unpaired t-tests with Benjamini–Hochberg FDR adjustment for multiple comparisons. NG2: P = 0.0018;

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: Expressing, Immunofluorescence, Quantitative RT-PCR, Control