Journal: bioRxiv

Article Title: NG2 glia protect against prion neurotoxicity by inhibiting prostaglandin E2 signaling

doi: 10.1101/2023.04.04.535590

Figure Lengend Snippet: A-B , NeuN immunofluorescence (A) and quantification (B) showing enhanced neurodegeneration in PGE2 treated compared to DMSO treated and PGD2 treated COCS after prion infection. n = 18 slices/condition. C-D , NeuN immunofluorescence (C) and quantification (D) showing diminished neurodegeneration in COCS treated with Ptges inhibitors C118 and C934 compared to DMSO treated COCS after prion infection. n = 18 slices/condition. E , Survival curves showing decelerated prion disease in Cox2 knockout (Cox2Luc) mice compared to littermate WT mice. F-G , Map2 immunofluorescence (F) and quantification (G) showing diminished dendritic pathology in the hippocampi of Cox2Luc mice after prion inoculation compared to littermate WT mice. Brain samples were collected at 21 wpi. n = 6 mice/group.

Article Snippet: Lenti-vectors used for expressing human PGE2 receptors EP1 (pLenti-PTGER1-mGFP-P2A- Puro, RC208597L4), EP2 (pLenti-PTGER2-mGFP-P2A-Puro, RC210883L4), EP3 (pLenti- PTGER3-mGFP-P2A-Puro, RC220173L4) and EP4 (pLenti-PTGER4-mGFP-P2A-Puro, RC210932L4) were purchased from OriGene Technologies.

Techniques: Immunofluorescence, Infection, Knock-Out

Journal: bioRxiv

Article Title: NG2 glia protect against prion neurotoxicity by inhibiting prostaglandin E2 signaling

doi: 10.1101/2023.04.04.535590

Figure Lengend Snippet: A-B , NeuN immunofluorescence (A) and quantification (B) showing that the enhanced neurodegeneration in prion-infected, NG2 glia-depleted (CP673451) COCS can be rescued by treatment with Ptges inhibitors C118 and C934. n = 16 slices/condition. C , Cox2 ablation (Cox2Luc) suppresses the acceleration of prion disease in NG2 glia-depleted (Pdgfrα iDTR ) mice. NG2 glia depletion was induced at 16 wpi. D-E , Map2 immunofluorescence (D) and quantification (E) showing enhanced dendritic pathology in NG2 glia-depleted (Pdgfrα iDTR ) hippocampi of prion-infected mice, and its rescue by Cox2 ablation (Cox2Luc). n = 6 mice/group. F-G , Live-cell images (F) and quantitative analysis (G) of chronically prion infected HovS cells expressing control (Ctrl) transgene or one of the four PGE2 receptors (Ptger). 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; 4 technical repeats/experiment

Article Snippet: Lenti-vectors used for expressing human PGE2 receptors EP1 (pLenti-PTGER1-mGFP-P2A- Puro, RC208597L4), EP2 (pLenti-PTGER2-mGFP-P2A-Puro, RC210883L4), EP3 (pLenti- PTGER3-mGFP-P2A-Puro, RC220173L4) and EP4 (pLenti-PTGER4-mGFP-P2A-Puro, RC210932L4) were purchased from OriGene Technologies.

Techniques: Immunofluorescence, Infection, Expressing, Control

Journal: Cell Reports Medicine

Article Title: Cross-Sectional Evaluation of Humoral Responses against SARS-CoV-2 Spike

doi: 10.1016/j.xcrm.2020.100126

Figure Lengend Snippet: Anti-S Neutralizing Antibody Titers Decrease over Time Pseudoviral particles coding for the luciferase reporter gene and bearing the glycoproteins SARS-CoV-2 S (A, D, G, and H), SARS-CoV S (B, E, and I), or VSV-G (C and F) were used to infect 293T-ACE2 cells. Pseudoviruses were incubated with serial dilutions of samples from 10 COVID-19-negative or 106 COVID-19-positive patients (T1, T2, T3, T4, and convalescent) at 37°C for 1 h prior to infection of 293T-ACE2 cells. Infectivity at each dilution was assessed in duplicate and is shown as the percentage of infection without sera for each glycoprotein. Neutralization half maximal inhibitory serum dilution (ID 50 ) (G and I) and ID 80 (H) values were determined using a normalized non-linear regression using GraphPad Prism software. Undetectable measures are represented as white symbols. Neutralizer represent patients with an ID 50 over 100 (G and I) or an ID 80 (H). Statistical significance was tested using Mann-Whitney U tests (∗p < 0.05; ∗∗p < 0.01).

Article Snippet: Briefly, 293T cells were co-transfected with two packaging plasmids (pLP1 and pLP2), an envelope plasmid (pSVCMV-IN-VSV-G) and a lentiviral transfer plasmid coding for human ACE2 (pLenti-C-mGFP-P2A-Puro-ACE2) (OriGene).

Techniques: Luciferase, Incubation, Infection, Neutralization, Software, MANN-WHITNEY

Journal: Cell Reports Medicine

Article Title: Cross-Sectional Evaluation of Humoral Responses against SARS-CoV-2 Spike

doi: 10.1016/j.xcrm.2020.100126

Figure Lengend Snippet:

Article Snippet: Briefly, 293T cells were co-transfected with two packaging plasmids (pLP1 and pLP2), an envelope plasmid (pSVCMV-IN-VSV-G) and a lentiviral transfer plasmid coding for human ACE2 (pLenti-C-mGFP-P2A-Puro-ACE2) (OriGene).

Techniques: Recombinant, Modification, Saline, Western Blot, Lysis, Expressing, Transfection, Staining, Labeling, Plasmid Preparation, Software, Flow Cytometry