Journal: PLoS ONE

Article Title: Aberrant Immune Responses in a Mouse with Behavioral Disorders

doi: 10.1371/journal.pone.0020912

Figure Lengend Snippet: Sera from postnatal day 21 mice were added to wells coated with BALB/c SCID whole brain proteins (10 µg/well). The numbers of sera were 6 for BTBR male or female, 6 for BCF1 male, 3 for BCF1 female, 5 for CBF1 male or female, and 7 for B6 male or female. The level of brain-reactive IgG was determined by using the HRP-conjugated goat anti-mouse IgG. *, p <0.01 vs. B6 males or females, and **, p <0.05 vs. B6 males.

Article Snippet: The level of total IgG in serum or brain homogenates was determined by a sandwich ELISA using goat anti-mouse IgG Fc (Pierce, Rockford, IL) as a capture Ab and HRP-goat anti-mouse IgG as a detection Ab (Sigma).

Techniques:

Journal: JCI Insight

Article Title: IL-13–programmed airway tuft cells produce PGE 2 , which promotes CFTR-dependent mucociliary function

doi: 10.1172/jci.insight.159832

Figure Lengend Snippet: ( A ) Among all DEGs for each cell type, 87 genes were upregulated in at least 9 cell types in polyp epithelium compared with controls and defined as pan-epithelial. ( B ) Fold change in normalized gene expression for tracheal epithelial cells cultured at air liquid interface (ALI) and stimulated with IL-13 ( n = 10 wells from 6 donors, * P < 0.05; ** P < 0.01 by ANOVA with Tukey correction). ( C ) Fold change in normalized SLC6A8 gene expression in human tracheal epithelial cells cultured at ALI and stimulated with indicated eicosanoids ( n = 9 donors, ** P < 0.01 by ANOVA with Dunnett’s correction). Represents similar responses for all genes as shown in D . ( D ) Fold change in normalized gene expression for tracheal epithelial cells cultured at ALI and stimulated with prostaglandin E 2 (PGE 2 ) ( n = 10 wells from 6 donors, ** P < 0.01; *** P < 0.001; **** P < 0.0001 by ANOVA with Tukey correction). ( E ) UMAP of scRNA-Seq data from surgical sinus tissue of participants with CRSwNP (“polyp”) or patients with CRS without nasal polyps (CRSsNP) (“no polyp”) whole polyp or nonpolyp sinus tissue . Epithelial clusters encircled with dashed line. ( F ) PGE 2 response gene score in polyp and nonpolyp epithelial clusters. * P < 0.05; ** P < 0.01; **** P < 0.0001 by linear mixed model. Statistical calculations relating to this figure are included in . For B and D , data shown as mean ± SEM. For C , horizontal line shows mean with bars indicating range.

Article Snippet: Overexpression of IL-13 was verified by measurement of serum IL-13 using a mouse IL-13 enhanced sensitivity flex set (BD), analyzed on an LSR Fortessa flow cytometer (BD) with FCAP Array software (BD) 1 week later, or at the time of sacrifice.

Techniques: Expressing, Cell Culture

Journal: JCI Insight

Article Title: IL-13–programmed airway tuft cells produce PGE 2 , which promotes CFTR-dependent mucociliary function

doi: 10.1172/jci.insight.159832

Figure Lengend Snippet: ( A ) T2 inflammatory mouse model system. ( B ) Representative sections of mouse nasal epithelium after 1 month of IL-13 overexpression or IgG control. Stars mark dual staining IL-25 + and DCLK1 + tuft cells. Bar indicates 50 μm. ( C ) Quantification of tuft cells in nasal epithelium in control and systemic IL-13 expression. ** P < 0.01 by t test. ( D ) Subclustering of tuft cells from control or IL-13–overexpressing mouse nasal epithelium. ( E ) Percentage of tuft cells derived from control or systemic IL-13 conditions in each subcluster in D . ( F ) Human polyp allergic tuft cell gene score in mouse nasal epithelial tuft cell subclusters. **** P < 0.0001 by linear regression model. ( G ) Schematic of protocol to measure ( H ) PGE 2 metabolites (PGEMs) in whole tracheal tissue from WT or Pou2f3 –/– mice exposed to systemic IL-13 or ( I ) PGE 2 in media from tracheal organoids derived from WT or Pou2f3 –/– mice. ( H and I ) * P < 0.05; *** P < 0.001 by t test. For C , H , and I , line is at mean and bars indicate max/min (range).

Article Snippet: Overexpression of IL-13 was verified by measurement of serum IL-13 using a mouse IL-13 enhanced sensitivity flex set (BD), analyzed on an LSR Fortessa flow cytometer (BD) with FCAP Array software (BD) 1 week later, or at the time of sacrifice.

Techniques: Over Expression, Control, Staining, Expressing, Derivative Assay