Journal: PLoS Pathogens

Article Title: Streptococcal M protein promotes IL-10 production by cGAS-independent activation of the STING signaling pathway

doi: 10.1371/journal.ppat.1006969

Figure Lengend Snippet: A) B6 macrophages were infected with M5 or ΔM5 bacteria, as indicated. Macrophages were lysed at the indicated time points (hours) post infection and analyzed for activation of Stat1 and Stat2 by Western blot. Uninfected (UI) macrophages were analyzed as control. Activated ( i . e . phosphorylated) transcription factors were detected with phospho-specific primary antibodies (indicated with p). Shown is one experiment representative of three. B) B6 macrophages were infected with M5 or ΔM5 for 4 hours, and IFNβ transcripts were measured by RTqPCR. mRNA levels are presented as fold-change relative to UI control macrophages. Results shown (mean and SD; n = 3 per group) are representative of three independent experiments. C) Kinetic analysis of IFNβ secretion from B6 macrophages infected as indicated. Results shown (mean; n = 2 per group) are representative of two independent experiments. D) B6 and IFNAR-KO macrophages were infected with M5 or ΔM5, or UI, as indicated. Culture supernatants were collected 24 hpi and assayed for indicated cytokines. Results shown (mean and SD; n = 3 per group) are representative of three independent experiments. E) B6 macrophages were infected with wild type M5 bacteria in the presence of titrated amounts (final concentration indicated) of a neutralizing anti-mouse IFNAR mAb or mouse IgG1 isotype control, as indicated. UI macrophages treated with the anti-IFNAR mAb were analyzed as control. Cytokines were assayed 24 hpi. Results shown (mean and SD; n = 3 per group) are representative of two independent experiments. F) B6 and IFNAR-KO macrophages were infected with S . pyogenes strains of six different serotypes (two that are serum opacity factor positive [OF + ], and four that are OF - ), as indicated. Cytokines were assayed 24 hpi. Results shown (mean and SD; n = 3 per group) are representative of two independent experiments. ANOVA (*<0.033; **<0.002; ***<0.001).

Article Snippet: Rabbit antibodies against mouse phospho-Stat2 (p-Tyr689) and Stat2 were from Millipore.

Techniques: Infection, Activation Assay, Western Blot, Concentration Assay

Journal: Infection and Immunity

Article Title: Legionella pneumophila inhibits type I interferon signaling to avoid cell-intrinsic host cell defense

doi: 10.1128/iai.00365-23

Figure Lengend Snippet: L. pneumophila inhibits cell surface expression of tetherin but does not affect the phosphorylation of STAT1 and STAT2. THP-1 cells were infected with GFP-expressing wild-type L. pneumophila and dotA deficient mutant (Lp∆) at an MOI of 10 and treated or not with 10 ng/mL IFN-β. (A) The flow cytometry analysis performed on the GFP+/infected cells using a fluorescent antibody targeting the cell surface ISG protein tetherin at 20 h.p.i.. (B) The flow cytometry analysis performed on the GFP+/infected cells at an MOI of 10 using a fluorescent antibody targeting phosphorylated STAT1 30 minutes post-infection. (C) The flow cytometry analysis performed on the GFP+/infected cells at an MOI of 10 using a fluorescent antibody targeting phosphorylated STAT2 30 minutes post-infection. The histograms are from one experiment representative of three independent experiments. The bar graphs show the mean fluorescent intensity fold changes compared with the uninfected untreated cells for each flow cytometry analysis. Individual points represent independent experiments. An asterisk indicates multiple unpaired t-tests with Welch correction relative to the uninfected IFN-β treated cells. Significance is indicated as follows: **P ≤ 0.01, ***P ≤ 0.001, and ****P ≤ 0.0001.

Article Snippet: Next, 50 μL of the stain cocktail [1× Brillant Stain Buffer (BD Bioscience) and either 5 μL per sample of Alexa Flour 647 Anti-Total Stat1 (N-Terminus) (BD Biosciences) and 5 μL per sample of PE anti-STAT1 Phospho (Tyr701) (BioLegend) or 2 μL per sample of Stat2 (D9J7L) Rabbit mAb (PE) (Cell Signaling Technology) and 2 μL per sample of P-Stat2 (P-690) Rabbit mAb (Alexa 647) (Cell Signaling Technology)] was added to each sample.

Techniques: Expressing, Phospho-proteomics, Infection, Mutagenesis, Flow Cytometry

Journal: International journal of oncology

Article Title: Downregulation of microRNA-431 by human interferon-β inhibits viability of medulloblastoma and glioblastoma cells via upregulation of SOCS6.

doi: 10.3892/ijo.2014.2317

Figure Lengend Snippet: Figure 5. Effects of HuIFN-β and miR-431 on SOCS6 expression and JAK- STAT signaling pathway in medulloblastoma cells. Immunoblot of SOCS6, JAK1, STAT1, p-STAT1, STAT2, p-STAT2, total Akt, p-Akt (Ser473), total Erk1/2, and p-Erk1/2 protein in medulloblastoma cells treated with or without HuIFN-β (1x105 IU/ml) and pre-miR-431 treatment for 48 h. GAPDH was used as a loading control.

Article Snippet: The following antibodies were used: SOCS6, JAK1 and total Akt1/2/3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); and STAT1 p-STAT1, STAT2, p-STAT2, p-Akt (phosphoAkt, Ser473), p44/42 MAPK (Erk1/2), and phosphor-p44/42 MAPK (Erk1/2) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Western Blot, Control

Journal: International journal of oncology

Article Title: Downregulation of microRNA-431 by human interferon-β inhibits viability of medulloblastoma and glioblastoma cells via upregulation of SOCS6.

doi: 10.3892/ijo.2014.2317

Figure Lengend Snippet: Figure 6. Effects of HuIFN-β and miR-431 on SOCS6 expression and JAK- STAT signaling pathway in glioblastoma cells. Immunoblot of SOCS6, JAK1, STAT1, p-STAT1, STAT2, p-STAT2, total Akt, p-Akt (Ser473), total Erk1/2, and p-Erk1/2 protein in glioblastoma cells treated with or without HuIFN-β (1x105 IU/ml) and pre-miR-431 treatment for 48 h. GAPDH was used as a loading control.

Article Snippet: The following antibodies were used: SOCS6, JAK1 and total Akt1/2/3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); and STAT1 p-STAT1, STAT2, p-STAT2, p-Akt (phosphoAkt, Ser473), p44/42 MAPK (Erk1/2), and phosphor-p44/42 MAPK (Erk1/2) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Western Blot, Control

Journal: International journal of oncology

Article Title: Downregulation of microRNA-431 by human interferon-β inhibits viability of medulloblastoma and glioblastoma cells via upregulation of SOCS6.

doi: 10.3892/ijo.2014.2317

Figure Lengend Snippet: Figure 5. Effects of HuIFN-β and miR-431 on SOCS6 expression and JAK- STAT signaling pathway in medulloblastoma cells. Immunoblot of SOCS6, JAK1, STAT1, p-STAT1, STAT2, p-STAT2, total Akt, p-Akt (Ser473), total Erk1/2, and p-Erk1/2 protein in medulloblastoma cells treated with or without HuIFN-β (1x105 IU/ml) and pre-miR-431 treatment for 48 h. GAPDH was used as a loading control.

Article Snippet: The following antibodies were used: SOCS6, JAK1 and total Akt1/2/3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); and STAT1 p-STAT1, STAT2, p-STAT2, p-Akt (phosphoAkt, Ser473), p44/42 MAPK (Erk1/2), and phosphor-p44/42 MAPK (Erk1/2) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Western Blot, Control

Journal: International journal of oncology

Article Title: Downregulation of microRNA-431 by human interferon-β inhibits viability of medulloblastoma and glioblastoma cells via upregulation of SOCS6.

doi: 10.3892/ijo.2014.2317

Figure Lengend Snippet: Figure 6. Effects of HuIFN-β and miR-431 on SOCS6 expression and JAK- STAT signaling pathway in glioblastoma cells. Immunoblot of SOCS6, JAK1, STAT1, p-STAT1, STAT2, p-STAT2, total Akt, p-Akt (Ser473), total Erk1/2, and p-Erk1/2 protein in glioblastoma cells treated with or without HuIFN-β (1x105 IU/ml) and pre-miR-431 treatment for 48 h. GAPDH was used as a loading control.

Article Snippet: The following antibodies were used: SOCS6, JAK1 and total Akt1/2/3 (Santa Cruz Biotechnology, Santa Cruz, CA, USA); and STAT1 p-STAT1, STAT2, p-STAT2, p-Akt (phosphoAkt, Ser473), p44/42 MAPK (Erk1/2), and phosphor-p44/42 MAPK (Erk1/2) (Cell Signaling Technology, Danvers, MA, USA).

Techniques: Expressing, Western Blot, Control