af1523  (R&D Systems)

Journal: Science Advances

Article Title: Mitochondria complex III–generated superoxide is essential for IL-10 secretion in macrophages

doi: 10.1126/sciadv.adu4369

Figure Lengend Snippet: RNA-seq analysis of QPC-HET ( n = 5) and QPC-KO ( n = 5) BMDMs 18 hours post–IL-4 (10 ng/ml) stimulation or vehicle control by: ( A ) Principal components analysis (PCA), ( B ) heatmap of gene expression of the top 20 genes, and ( C ) summed module expression of the IL-4 response module as defined in . Frequency of ( D ) Arg-1 + , ( E ) Relm-α + , and ( F ) PD-L2 + BMDMs 18 hours post–IL-4 (10 ng/ml) stimulation. Frequency of ( G ) Arg-1 + and ( H ) Relm-α + peritoneal macrophages 4 days post-initial thioglycolate and IL-4c stimulation QPC-HET (unstimulated, n = 3; IL-4c stimulated, n = 10) and QPC-KO (unstimulated n = 3; IL-4c stimulated, n = 12). Data represent means ± SEM. In (A), PCA was performed using single value decomposition of the top 1000 genes by variance. In (B), hierarchical clustering was performed using Ward’s method (D2) with Euclidean distance as a distance metric. Gene expression values are z -normalized DESeq2 counts. In (C), pairwise comparisons were performed using pairwise Mann-Whitney U tests with FDR correction. In (D) to (H), ordinary one-way ANOVAs with Holm-Sidak’s multiple comparisons test with a single pooled variance were used to compare groups. Data represent means ± SEM.

Article Snippet: Fixation and permeabilization were performed with BD Fix/Perm (BD, 51-2090KZ) and, subsequently, stained with 5.5 μl of anti-RELMα (PerCP-efluor710, Invitrogen, 53-3697-82, clone DS8RELM), 5.5 μl of anti–Arg-1 (FITC, Invitrogen, 53-3697-82, clone: A1exF5), and anti-CD206 (BV421, BioLegend, 141717, clone C068C2).

Techniques: RNA Sequencing Assay, Control, Expressing, MANN-WHITNEY

Journal: Science Advances

Article Title: Mitochondria complex III–generated superoxide is essential for IL-10 secretion in macrophages

doi: 10.1126/sciadv.adu4369

Figure Lengend Snippet: RNA-seq analysis of QPC-HET ( n = 5) and QPC-KO ( n = 5) BMDMs 18 hours post–IL-4 (10 ng/ml) stimulation or vehicle control by: ( A ) Principal components analysis (PCA), ( B ) heatmap of gene expression of the top 20 genes, and ( C ) summed module expression of the IL-4 response module as defined in . Frequency of ( D ) Arg-1 + , ( E ) Relm-α + , and ( F ) PD-L2 + BMDMs 18 hours post–IL-4 (10 ng/ml) stimulation. Frequency of ( G ) Arg-1 + and ( H ) Relm-α + peritoneal macrophages 4 days post-initial thioglycolate and IL-4c stimulation QPC-HET (unstimulated, n = 3; IL-4c stimulated, n = 10) and QPC-KO (unstimulated n = 3; IL-4c stimulated, n = 12). Data represent means ± SEM. In (A), PCA was performed using single value decomposition of the top 1000 genes by variance. In (B), hierarchical clustering was performed using Ward’s method (D2) with Euclidean distance as a distance metric. Gene expression values are z -normalized DESeq2 counts. In (C), pairwise comparisons were performed using pairwise Mann-Whitney U tests with FDR correction. In (D) to (H), ordinary one-way ANOVAs with Holm-Sidak’s multiple comparisons test with a single pooled variance were used to compare groups. Data represent means ± SEM.

Article Snippet: Fixation and permeabilization were performed with BD Fix/Perm (BD, 51-2090KZ) and, subsequently, stained with 5.5 μl of anti-RELMα (APC, Invitrogen, 17-5441-82, clone: DS8RELM) and 5.5 μl of anti–Arg-1 [fluorescein isothiocyanate (FITC), Intivrogen, 53-3697-82, clone: A1exF5].

Techniques: RNA Sequencing Assay, Control, Expressing, MANN-WHITNEY