Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Loss of Mitochondrial Protease CLPP Activates Type I IFN Responses through the Mitochondrial DNA–cGAS–STING Signaling Axis

doi: 10.4049/jimmunol.2001016

Figure Lengend Snippet: (A–D) WT and CLPP-KO MEFs were cultured in the presence of regular media (Control) or media containing ddC (150μM). Cells were harvested after 4 days for (A) confocal microscopy with anti-DNA (DNA), anti-Tfam (mtDNA nucleoid marker) and anti-HSP60 (mitochondrial matrix protein). (B) Nucleoid quantification represented as percentage (%) of enlarged nucleoids (>450nm2). (C) Quantitative real-time PCR analysis of ISG expression (n=3 biological replicates). (D) Representative western blots of ISG protein expression. (E) Quantitative real-time PCR of ISGs in CLPP-KO MEFs transfected with control or siPolg2 siRNA for 72hrs. (n=3 biological replicates). (F) Quantitative real-time PCR of ISGs of WT and CLPP-KO MEFs treated with VBIT-4 (10μM) or Control (DMSO) for 48hrs (n=3 biological replicates). In (E), real-time PCR data are presented as relative expression percentage (%) setting siControl as 100%. Error bars represent mean ± s.e.m. of triplicates, Student’s t-test. In (C) and (F), error bars represent mean ± s.e.m. of triplicates, two-way ANOVA Tukey’s post-hoc. *p<0.05, **p<0.01, ****p<0.0001. All experiments are representative of 3–4 independent experiments.

Article Snippet: For VBIT-4 treatment plus challenge, cells were treated with VBIT-4 and 6hrs before the 48hrs harvesting time. mIFNβ (8234-MB-010/CF, R&D) was added to the cells at a final concentration of 1ng/ml.

Techniques: Cell Culture, Confocal Microscopy, Marker, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Transfection

Journal: Nature communications

Article Title: Comparative therapeutic efficacy of remdesivir and combination lopinavir, ritonavir, and interferon beta against MERS-CoV.

doi: 10.1038/s41467-019-13940-6

Figure Lengend Snippet: Fig. 5 Therapeutic RDV reduces replication and pathology. Percent starting weight of 10–12-week-old female Ces1c−/−hDPP4 mice infected with 5E + 04 pfu MERS M35C4 and treated with a subcutaneous vehicle for RDV (N = 13) or remdesivir (RDV, 25 mg/kg, N = 14) BID beginning 1 dpi or b vehicle for LPV/RTV-IFNb (N = 15), LPV/RTV-IFNb low (N = 16) or LPV/RTV-IFNb high (N = 16) beginning 1 dpi. Oral vehicle or lopinavir/ritonavir (160/40 mg/kg) was administered orally once daily. IFNb low (1x human equivalent dose of 1.6 MIU/kg) and high (25x human equivalent dose of 40 MIU/kg) or PBS vehicle were administered via subcutaneous injection every other day. Asterisks indicate statistical differences by two-way ANOVA with Tukey’s multiple comparison test. c Lung hemorrhage 6 dpi for all animals in a, b scored on a scale of 0–4, where 0 is a normal pink healthy lung and 4 is a diffusely discolored dark red lung. d MERS-CoV lung titer 6 dpi in mice as described in a, b. Asterisks indicate statistical significance (N group described in a and b, P < 0.05) by one-way ANOVA with Kruskal–Wallis test for (c, d). Data for a–d are compiled from two independent experiments. For the box and whisker plots, the boxes encompass the 25th to 75th percentile, the line is at the median, while the whiskers represent the range. e Representative photomicrographs of MERS-CoV antigen (brown) and hematoxylin stained nuclei (blue) in mouse lung tissue sections from 6 dpi. The black bar is 100 µM.

Article Snippet: Recombinant mouse IFNb protein was purchased from R&D Systems (8234-MB/CF, 1.2 × 109 IU/mg calibrated against Murine IFN-beta WHO International Standard) for the in vivo studies and reconstituted in PBS.

Techniques: Infection, Injection, Comparison, Whisker Assay, Staining