Journal: EBioMedicine

Article Title: Therapeutic efficacy of pulmonary live tuberculosis vaccines against established asthma by subverting local immune environment

doi: 10.1016/j.ebiom.2020.103186

Figure Lengend Snippet: BCG and MTBVAC intranasal revert established allergic airway responsiveness in an OVA-driven chronic model. (a) Scheme of the experiment based on an OVA-induced chronic model. Sensitized mice were challenged from week 3 to 13 with two intranasal inoculations weekly of OVA 10μg. Vaccines were delivered at week 9, in the half of the challenge phase. (b) Total number of eosinophils in BAL at week 8, the week before vaccine immunization. (c) Total number of eosinophils in BAL at week 13, four weeks after BCG treatment. (d) Total number of eosinophils in BAL at week 13, four weeks after treatment with 10 7 CFU of MTBVAC. (e) Representative images of PAS-stained fixed lungs from OVA-challenged mice untreated or MTBVAC-treated. (f, g) Intracellular expression of iNOS and Arg-1 in lung macrophages at week 13. Representative dot-plot diagrams are shown. Data in the graphs are pooled means±SEM from two independent experiments (b, c), or one (d, g). A minimum of 6 mice was used per group and experiment. * p <0.05; ** p <0.01; *** p <0.001, by t-student test (b, e), one-way ANOVA with Bonferroni post-test (b, c), and multiple t-student test (g).

Article Snippet: As intracellular antibodies, we used iNOS-APC (RRID:AB_2727527) and iNOS-PE (RRID:AB_2727486) (Miltenyi), and Arg1-APC (RRID:AB_2734835)(eBiosciences).

Techniques: Vaccines, Staining, Expressing

Journal: EBioMedicine

Article Title: Therapeutic efficacy of pulmonary live tuberculosis vaccines against established asthma by subverting local immune environment

doi: 10.1016/j.ebiom.2020.103186

Figure Lengend Snippet:

Article Snippet: As intracellular antibodies, we used iNOS-APC (RRID:AB_2727527) and iNOS-PE (RRID:AB_2727486) (Miltenyi), and Arg1-APC (RRID:AB_2734835)(eBiosciences).

Techniques: Sequencing

Journal: Cell Communication and Signaling : CCS

Article Title: A specific, non-immune system-related isoform of the human inducible nitric oxide synthase is expressed during differentiation of human stem cells into various cell types

doi: 10.1186/s12964-022-00855-x

Figure Lengend Snippet: NOS2 transcripts in the ENSEMBL database

Article Snippet: In addition to NOS2 expression in the different colon parts, the human protein atlas describes NOS2 expression in human placental cells.

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: A specific, non-immune system-related isoform of the human inducible nitric oxide synthase is expressed during differentiation of human stem cells into various cell types

doi: 10.1186/s12964-022-00855-x

Figure Lengend Snippet: NOS2 mRNA isoform expression in the human sigmoid colon and small intestine. RNA-Seq data (gene expression omnibus GSM1010942, GSM1010940) were analyzed for human NOS2 mRNA isoform expression. Shown are the mean ± SEM of the rpkm values of the different NOS2 mRNA isoforms (NOS2-1, -2 and -3). There was no expression of NOS2-2 and -3

Article Snippet: In addition to NOS2 expression in the different colon parts, the human protein atlas describes NOS2 expression in human placental cells.

Techniques: Expressing, RNA Sequencing, Gene Expression

Journal: Cell Communication and Signaling : CCS

Article Title: A specific, non-immune system-related isoform of the human inducible nitric oxide synthase is expressed during differentiation of human stem cells into various cell types

doi: 10.1186/s12964-022-00855-x

Figure Lengend Snippet: NOS2 mRNA isoform expression in isolated human islets treated with or without a cytokine mixture. RNA-Seq data (Bioproject PRJNA151601 ) were analyzed for human NOS2 mRNA isoform expression. Shown are the mean ± SEM of the rpkm values of the different NOS2 mRNA isoforms (NOS2-1, -2 and -3) expressed in isolated human islets incubated with (CM) or without a cytokine mixture (co) containing IL-1β and IFN-γ. ***FDR p value < 0.001, ns FDR p value > 0.05 versus co

Article Snippet: In addition to NOS2 expression in the different colon parts, the human protein atlas describes NOS2 expression in human placental cells.

Techniques: Expressing, Isolation, RNA Sequencing, Incubation

Journal: Cell Communication and Signaling : CCS

Article Title: A specific, non-immune system-related isoform of the human inducible nitric oxide synthase is expressed during differentiation of human stem cells into various cell types

doi: 10.1186/s12964-022-00855-x

Figure Lengend Snippet: NOS2 mRNA isoform expression in different placenta-derived trophoblast stem cells (CT29, CT30) and H1- or H9-ESC induced to differentiate to trophoblast stem cells. RNA-Seq data (Bioproject PRJNA565033 were analyzed for human NOS2 mRNA isoform expression. Shown are the mean ± SEM of the rpkm values of NOS2 mRNA isoforms at different differentiation stages. ***FDR p value < 0.001, ns FDR-value > 0.05 versus untreated H9-ESC

Article Snippet: In addition to NOS2 expression in the different colon parts, the human protein atlas describes NOS2 expression in human placental cells.

Techniques: Expressing, Derivative Assay, RNA Sequencing

Journal: Cell Communication and Signaling : CCS

Article Title: A specific, non-immune system-related isoform of the human inducible nitric oxide synthase is expressed during differentiation of human stem cells into various cell types

doi: 10.1186/s12964-022-00855-x

Figure Lengend Snippet: NOS2 mRNA isoform expression in human iPSC from normal (WT) and down syndrome (DOWN) donors induced to differentiate to neurons . RNA-Seq data (Bioproject PRJDB1099 ; FANTOM5) were analyzed for human NOS2 mRNA expression. Shown are the mean ± SEM of the rpkm values of the different NOS2 mRNA isoforms (NOS2-1, -2 and -3) at different time points (day 0 to day 18). ***FDR p value < 0.001, *FDR p value < 0.05, ns FDR p value > 0.05 versus day 0

Article Snippet: In addition to NOS2 expression in the different colon parts, the human protein atlas describes NOS2 expression in human placental cells.

Techniques: Expressing, RNA Sequencing

Journal: Cell Communication and Signaling : CCS

Article Title: A specific, non-immune system-related isoform of the human inducible nitric oxide synthase is expressed during differentiation of human stem cells into various cell types

doi: 10.1186/s12964-022-00855-x

Figure Lengend Snippet: NOS2 mRNA- and protein expression in four different human iPSC lines induced to differentiate to neurons. Four different human iPSCs lines were generated from PBMC of three different donors . RNA and protein were isolated from these 4 different human iPSC lines induced to differentiate to neurons at different time points (day 0 to 60). A NOS2-mRNA and 18S rRNA expression were analyzed using the qRT-PCR method. NOS2 mRNA expression was normalized to the 18S rRNA expression. The relative NOS2 mRNA expression in the cells treated for 18 days was set to 100%. Shown is the summary of the analysis of the four different iPSC cell lines. The values represent the mean ± SEM of n = 12 different isolated RNAs at each time point. (*** p < 0.001, * p < 0.05, ns not significant vs. iPSC treated for zero days; 1-way Anova with Dunnett's multiple comparisons test). B NOS2- and GAPDH protein expression in DLD1 cells (DLD1) incubated with (CM) or without (Co) a cytokine mixture to induce the human iNOS expression and ILB-C89bf (ILB-C89bf 18 days) cells stimulated to differentiate to neurons for 18 days were analyzed using the western blot method. Shown are the detected bands of NOS2- and GAPDH protein (analyzed using either the anti-NOS2- or the anti-GAPDH antibody)

Article Snippet: In addition to NOS2 expression in the different colon parts, the human protein atlas describes NOS2 expression in human placental cells.

Techniques: Expressing, Generated, Isolation, Quantitative RT-PCR, Incubation, Western Blot

Journal: Cell Communication and Signaling : CCS

Article Title: A specific, non-immune system-related isoform of the human inducible nitric oxide synthase is expressed during differentiation of human stem cells into various cell types

doi: 10.1186/s12964-022-00855-x

Figure Lengend Snippet: NOS2 mRNA isoform analysis using 5′RACE and isoform specific qRT-PCR methods. A Schematic summary of the results of 5′-RACE analyses using RNA isolated at day 18 from human iPSC induced to differentiate to neurons. RNA was isolated from 4 different human iPSC lines (samples C16-1, C16-2, C89-2, C89-3) induced to differentiate to neurons for 18 days. As control RNA was isolated from human DLD1 cells (DLD1-2 and DLD1-2) treated with a cytokine mixture for 6 h to induce NOS2 mRNA expression. Then 5′-RACE experiments were performed (as described in “Methods” section) using the indicated oligonucleotides as 3′-primer (5′RACE-rev = primer used for the RT-reaction; 3P2/3P3 primers used for the PCR-reaction). The final PCR products were isolated, cloned into pCR-Script and sequenced. The alignment of these sequences to the human NOS2-1 and -2 mRNA are indicated as filled arrows. The sequences of the fragments are shown in Additional file : Fig. S10. B/C NOS2-mRNA-isoform expression analysis using isoform specific qRT-PCR experiments. RNA was isolated at different time points from 2 different human iPSC lines (iLB-C16bm s6 iPS andiLB-C89bf s4 iPCs) induced to differentiate to neurons for 60 days. The RT reaction was performed with the RT-rev primer (see A ). Taqman qPCR reactions were performed with mRNA isoform specific primer pairs and taqman probes (NOS2-1: NOS2-1_P, NOS2-1_3P and NOS2-1_Taq; NOS2-2: NOS2-2_5P, NOS2-2_3P and NOS2-2_Taq—see A ). For normalization also the GAPDH mRNA expression was measured. B The normalized NOS2-1 mRNA expression values were related to the CM induced NOS2-1 mRNA expression in DLD-1 cells (CM = 100%). Also, the expression level of the NOS2-1 mRNA in untreated (co) DLD-1 cells were determined. (*** p < 0.001, ns not significant vs. iPSC treated for 18 days; 1-way Anova with Dunnett’s multiple comparisons test). C The normalized NOS2-2 mRNA expression values were related to the NOS2-2 mRNA expression in iPSC treated for 18 days (day 18 = 100%). (** p < 0.05, vs. iPSC treated for 18 days; 1-way Anova with Dunnett’s multiple comparisons test)

Article Snippet: In addition to NOS2 expression in the different colon parts, the human protein atlas describes NOS2 expression in human placental cells.

Techniques: Quantitative RT-PCR, Isolation, Control, Expressing, Clone Assay

Journal: Cell Communication and Signaling : CCS

Article Title: A specific, non-immune system-related isoform of the human inducible nitric oxide synthase is expressed during differentiation of human stem cells into various cell types

doi: 10.1186/s12964-022-00855-x

Figure Lengend Snippet: The NOS2-2 protein is functional. DLD-1_TR7 cells stably expressing a tetracycline repressor were transiently transfected with pcDNA4/TO_NOS2-1_cds_3UTR or pcDNA4/TO_NOS2-2_cds_3UTR encoding the NOS2-1 or -2 protein. To normalize the transfection efficiency, pRL-EF1α (encoding for a renilla luciferase) was cotransfected as well. After transfection, the cells were incubated with (Tet) or without (co) 500 ng/ml tetracycline for 24 h. Then the supernatants of the cells were used for nitrate concentration determination by the Griess assay. The cells were lyzed and renilla luciferase activity was measured. The nitrate concentrations determined were normalized to the renilla luciferase data. Shown is the summary of the analysis of the four different transfection experiments. The values represent the mean ± SEM of n = 8 different wells, (*** p < 0.001; * p < 0.05, ns not significant;1-way Anova with Dunnett's multiple comparisons test)

Article Snippet: In addition to NOS2 expression in the different colon parts, the human protein atlas describes NOS2 expression in human placental cells.

Techniques: Functional Assay, Stable Transfection, Expressing, Transfection, Luciferase, Incubation, Concentration Assay, Griess Assay, Activity Assay

Journal: Cell Communication and Signaling : CCS

Article Title: A specific, non-immune system-related isoform of the human inducible nitric oxide synthase is expressed during differentiation of human stem cells into various cell types

doi: 10.1186/s12964-022-00855-x

Figure Lengend Snippet:

Article Snippet: In addition to NOS2 expression in the different colon parts, the human protein atlas describes NOS2 expression in human placental cells.

Techniques: