Journal: The American Journal of Pathology

Article Title: A Protein C Deficiency Exacerbates Inflammatory and Hypotensive Responses in Mice During Polymicrobial Sepsis in a Cecal Ligation and Puncture Model

doi:

Figure Lengend Snippet: Plasma coagulation factor assays of WT and PC+/− mice after CLP surgery. a: Plasma PC levels. The plasma PC levels were determined by ELISA before (WT, n = 4; PC+/−, n = 5), and 6 hours (WT, n = 5; PC+/−, n = 7) and 24 hours (WT, n = 5; PC+/−, n = 5) after CLP surgery in WT and PC+/− male mice. *, P < 0.001; #, P < 0.01. b: Plasma fibrinogen levels were determined by a coagulometric assay at 6 hours (WT, n = 4; PC+/−, n = 5) and 24 hours (WT, n = 15; PC+/−, n = 14) after, CLP surgery and in resting (WT, n = 11; PC+/−, n = 9) WT and PC+/− male mice. *, P = 0.016. c: Plasma PAI-1 levels were determined by ELISA before (WT, n = 3; PC+/−, n = 4), and 6 hours (WT, n = 3; PC+/−, n = 7) and 24 hours (WT, n = 5; PC+/−, n = 4) after, CLP surgery in WT and PC+/− male mice. d: Plasma FXII levels relative to the resting WT group, as measured by a coagulometric assay, before (WT, n = 4; PC+/−, n = 3) and 24 hours after (WT, n = 5; PC+/−, n = 3), CLP surgery. *, P = 0.05 (Kruskal-Wallis). e: Plasma FXI levels relative to the resting WT group as measured by a coagulometric assay, before (WT, n = 7; PC+/−, n = 5) and 24 hours after (WT, n = 5; PC+/−, n = 3), CLP surgery. No significant differences were observed.

Article Snippet: After four washes with PBS/Tween-20, 100 μl of 4 μg/ml rabbit anti-mouse PAI-1 polyclonal antibody (Molecular Innovations) in PBS was added to the wells and incubated for 2 hours.

Techniques: Coagulation, Enzyme-linked Immunosorbent Assay

Journal: The American Journal of Pathology

Article Title: A Protein C Deficiency Exacerbates Inflammatory and Hypotensive Responses in Mice During Polymicrobial Sepsis in a Cecal Ligation and Puncture Model

doi:

Figure Lengend Snippet: Gene expression ratio to RPL-19 of PAI-1 (a), MPO (b), IL-1β (c), and IL-6 (d) relative to the WT resting group (n = 3) in liver, kidney, and lung, as determined by RT-PCR in WT mice 24 hours after CLP (n = 3), PC+/− resting mice (n = 3), and PC+/− mice 24 hours after CLP (n = 4). a: In liver, PC+/− mice 24 hours after CLP were different from PC+/− resting and WT resting groups (P = 0.025, K-W), in kidney, differences between groups were not significant (K-W, multicomparison test). b: In liver, PC+/− resting mice were different from PC+/− mice 24 hours after CLP and WT resting groups (P = 0.055); in kidney, PC+/− mice 24 hours after CLP were different from all of the other groups (P = 0.02, K-W); in lung, PC+/− resting mice were different from PC+/− and WT mice 24 hours after CLP (P = 0.08, K-W). c: In liver, no significant differences between groups were found; in kidney: PC+/− group 24 hours after CLP was different from WT and PC+/− resting groups (P = 0.04, K-W); in lung: the PC+/− group after CLP was not different form the PC+/− resting group (T-K); P = not significant (K-W). d: In liver, PC+/− mice 24 hours after CLP were different from the WT resting group (P = 0.11, K-W); in kidney, PC+/− mice 24 hours after CLP were different from the WT and PC+/− resting groups (P = 0.032, K-W); in lung: no significant differences between the groups were found (K-W).

Article Snippet: After four washes with PBS/Tween-20, 100 μl of 4 μg/ml rabbit anti-mouse PAI-1 polyclonal antibody (Molecular Innovations) in PBS was added to the wells and incubated for 2 hours.

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

Journal: bioRxiv

Article Title: A Highly Immunogenic Measles Virus-based Th1-biased COVID-19 Vaccine

doi: 10.1101/2020.07.11.198291

Figure Lengend Snippet: (A) Schematic depiction of full-length SARS-CoV-2 S and recombinant MeV vac2 genomes used for expression of this antigen (lower schemes). Antigen or antigen encoding genes are depicted in dark grey; MeV viral gene cassettes (in light grey) are annotated. MluI and AatII restriction sites used for cloning of antigen-genes into post P or post H ATU are highlighted (B) Immunoblot analysis of Vero cells infected at an MOI of 0.01 with MeV vac2 -SARS2-S(P), MeV vac2 -SARS2-S(H), or MV vac2 -ATU(P) (MV vac2 ) as depicted above lanes. Uninfected cells served as mock. Blots were probed using rabbit polyclonal anti-SARS spike antibody (upper blot) or mAb reactive against MeV-N (lower blot). Arrows indicate specific bands. ( C, D ) Growth kinetics of recombinant MeV on Vero cells infected at an MOI of 0.03 with MV vac2 -ATU(P) or MeV vac2 -SARS2-S encoding extra genes in post H or post P. Titers of samples prepared at indicated time points post infection were titrated on Vero cells. Means and standard deviations of three to five independent experiments are presented. ( E ) SARS-CoV-2 S protein expression in Vero cells was verified via immunoperoxidase monolayer assay. 50× magnification; scale bar, 500 μm.

Article Snippet: Donkey anti-rabbit IgG-HRP (H&L) polyclonal antibody (1:10,000; 611-7202; Rockland) and goat anti-mouse IgG-HRP (1:10,000; A2554-1ML; Merck, Darmstadt, Germany) served as secondary antibodies.

Techniques: Recombinant, Expressing, Clone Assay, Western Blot, Infection