p28 subunit  (Cusabio)

Journal: International Journal of Molecular Sciences

Article Title: Myeloid-Derived Suppressor Cells Gain Suppressive Function during Neonatal Bacterial Sepsis

doi: 10.3390/ijms22137047

Figure Lengend Snippet: Changes in the MDSC effector gene expression during an infection. C57BL/6 neonatal pups were infected on day 3 or 4 of life with ~10 6 CFUs. At 18–24 h post-infection, the spleens were harvested from the infected or control mice and MDSCs were isolated by an immunomagnetic selection. RNA was isolated and the expression of nitric oxide synthase (NOS)2, arginase (ARG)1, catalytic subunit of the NADPH oxidase complex (CybB), interleukin (IL)-27p28 and Epstein-Barr virus induced 3 (EBI3) genes was measured by quantitative real time PCR. The values were normalized to β-actin and expressed as a log 2 change in expression relative to the uninfected control using the formula 2 −ΔΔCt . The mean results ± standard error from four experiments for NOS2 and three experiments for ARG1, CybB, p28 and EBI3 are shown. An asterisk indicates a statistical significance in the 95% confidence interval relative to the uninfected control that was determined using a paired sample t -test.

Article Snippet: Interleukin-27 subunit p28 (IL-27p28) , ThermoFisher , Mm00461164_m1.

Techniques: Gene Expression, Infection, Control, Isolation, Selection, Expressing, Virus, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Myeloid-Derived Suppressor Cells Gain Suppressive Function during Neonatal Bacterial Sepsis

doi: 10.3390/ijms22137047

Figure Lengend Snippet: MDSC change in function is directly regulated by bacteria. MDSCs were isolated from the spleens of neonatal naïve mice by an immunomagnetic selection and cultured with E. coli at a target MOI of 10 for 30 min prior to the addition of gentamicin (100 µg/mL). ( A ) RNA was isolated at 18–24 h post-infection and the expression of nitric oxide synthase (NOS)2, arginase (ARG)1, catalytic subunit of the NADPH oxidase complex (CybB), interleukin (IL)-27p28 and Epstein-Barr virus induced 3 (EBI3) genes was measured by quantitative real time PCR. The values were normalized to β-actin and expressed as a log 2 change in expression relative to the uninfected controls using the formula 2 −ΔΔCt . The mean results ± standard error from five to six experiments are shown. An asterisk indicates a statistical significance in the 95% confidence interval that was determined using a paired sample t -test. ( B ) Nitrite levels in the supernatants of the control or infected MDSCs were quantified as described in the Methods section. The mean nitrite levels ± standard error from four experiments are shown. ( C ) Arginase activity was measured from the cell lysates of MDSCs as described in the Methods section. The mean arginase levels ± standard error from six experiments are shown. ( D ) IL-27 concentrations in the supernatants of the control or infected MDSCs were measured by an ELISA. The mean concentrations ± standard error from three experiments are shown. An asterisk indicates a statistical significance in the 95% confidence interval that was determined using a Wilcoxon matched-pairs rank test.

Article Snippet: Interleukin-27 subunit p28 (IL-27p28) , ThermoFisher , Mm00461164_m1.

Techniques: Bacteria, Isolation, Selection, Cell Culture, Infection, Expressing, Virus, Real-time Polymerase Chain Reaction, Control, Activity Assay, Enzyme-linked Immunosorbent Assay

Journal: International Journal of Molecular Sciences

Article Title: Myeloid-Derived Suppressor Cells Gain Suppressive Function during Neonatal Bacterial Sepsis

doi: 10.3390/ijms22137047

Figure Lengend Snippet: Bacterial components elicit a variable stimulation of the effector gene expression compared with E. coli . MDSCs were isolated from the spleens of neonatal control mice by an immunomagnetic selection and cultured with E. coli at a target MOI of 10 for 30 min prior to the addition of gentamicin (100 µg/mL) or stimulated with lipopolysaccharide (LPS) (1 µg/mL), flagellin (100 ng/mL), peptidoglycan (PGN) (10 µg/mL), or a cocktail of each component at the described concentrations (combo). RNA was isolated 18–24 h post-infection or stimulation and the expression of NOS2, Arg-1 and IL-27p28 genes was measured by quantitative real time PCR. The values were normalized to β-actin and expressed as a log 2 change relative to the uninfected controls using the formula 2 −ΔΔCt . The mean expression ± standard error from seven experiments for individual bacterial components and three experiments for the combination of components are shown. An asterisk indicates a statistical significance in the 95% confidence interval that was determined using a two-way mixed model analysis of variance. Asterisks indicate a statistically significant difference in the expression between individual or the combination of components compared with E. coli within a gene.

Article Snippet: Interleukin-27 subunit p28 (IL-27p28) , ThermoFisher , Mm00461164_m1.

Techniques: Gene Expression, Isolation, Control, Selection, Cell Culture, Infection, Expressing, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Sciences

Article Title: Myeloid-Derived Suppressor Cells Gain Suppressive Function during Neonatal Bacterial Sepsis

doi: 10.3390/ijms22137047

Figure Lengend Snippet: Detailed primer information for those used in the gene expression studies.

Article Snippet: Interleukin-27 subunit p28 (IL-27p28) , ThermoFisher , Mm00461164_m1.

Techniques: Gene Expression, Sequencing