mouse il 1ra il 1f3 duoset kit  (R&D Systems)

Journal: PLOS Pathogens

Article Title: Single-cell transcriptomics unveils skin cell specific antifungal immune responses and IL-1Ra- IL-1R immune evasion strategies of emerging fungal pathogen Candida auris

doi: 10.1371/journal.ppat.1012699

Figure Lengend Snippet: (A) Heatmap represents the expression of Il1rn in the cell types (y-axis) between the infected and uninfected groups. The normalized gene counts were plotted in the heatmap, and the scale indicates red for high, blue for low, and white for moderate expression in the samples. Each column represents a different sample. (B) The violin plot depicts the scaled count of Il1rn in myeloid subsets from both uninfected and infected samples. The scaled count were normalized using SCTransform method. IL-1Ra levels in the (C) skin and (D) serum of mice after 3 DPI of C . auris AR0387 or C . albicans SC5314 ( n = 8–10 mice/group). Uninfected mice received 100 μL of 1X PBS on the day of infection. (E) Fungal burden in the skin tissue of indicated mice on 3 DPI of C . auris AR0387 or C . albicans SC5314 ( n = 8–10 mice/group). Uninfected mice received 100 μL of 1X PBS on the day of infection. (F) The bar graph represents the neutrophil-killing activity of C . auris AR0387 in the presence and absence of recombinant IL-1Ra. (G) Skin fungal burden of the mice groups injected with neutralizing anti-mouse IL-1Ra monoclonal antibody or 100 μl of 1X PBS after 3 DPI of C . auris AR0387 in the murine skin. ( n = 3–5 mice/group) (H-K) Quantification of IL-1Ra level and fungal burden in mice injected with anti-Ly6G antibody and Rat IgG2a isotype control (Isotype) for neutrophil depletion after 2 DPI of C . auris AR0387 ( n = 6–8 mice/group). H) Representative flow plots and I) percentage and absolute number of CD11b + Ly6G + neutrophils in the infected skin tissue of mice injected with anti-Ly6G antibody or Rat IgG2a isotype control (Isotype) antibody. (J) IL-1Ra level and (K) CFU were determined from the homogenate of infected skin tissue. (L-O) Measurement of IL-1Ra production and fungal burden in clodrosome (C.L) and 1X PBS (Ctrl) injected mice groups for macrophage depletion after 2 DPI of C . auris AR0387 ( n = 9–10 mice/group). (L) Representative flow plots of F4/80+ MHCII+ macrophage in the infected skin tissue of mice injected with clodrosome (C.L) or 1X PBS (Ctrl) injected mice groups. (M) IL-1Ra level and (N) CFU determined from the homogenate of infected skin tissue from 2 DPI of C . auris 0387 (n = 12 mice/group).(O) IL-1Ra level and (P) CFU determined from the homogenate of infected skin tissue from 12 DPI of C . auris 0387 (n = 8–10 mice/group). Error bars represent mean ± SEM. * p < 0.05, ** p <0.01, *** p <0.001, **** p <0.0001, NS, non-significant. Statistical significances were calculated using Mann–Whitney U. Abbreviations–DPI, days post-infection; C.L, clodrosome.

Article Snippet: IL-1Ra production in skin homogenate, culture supernatant, and serum was quantified using a mouse IL-1Ra/IL-1F3 DuoSet kit (RnD Systems) by following the manufacturer’s standard operating protocol.

Techniques: Expressing, Infection, Activity Assay, Recombinant, Injection, Control, MANN-WHITNEY

Journal: PLOS Pathogens

Article Title: Single-cell transcriptomics unveils skin cell specific antifungal immune responses and IL-1Ra- IL-1R immune evasion strategies of emerging fungal pathogen Candida auris

doi: 10.1371/journal.ppat.1012699

Figure Lengend Snippet: (A) The representative flow plots represents the percentage of CD11b + Ly6G + neutrophils and CD64 + MHCII + macrophage in the WT and Il1r1 -/- mice skin tissue infected with C . auris 0387. The bar graph represents the percentage and absolute number of CD11b + Ly6G + neutrophils after (B) 3 DPI and (C) 12 DPI and the CD64 + MHCII + macrophage after (D) 3 DPI and (E) 12 DPI in the WT and Il1r1 -/- mice skin tissue infected with C . auris 0387 ( n = 4–8 mice/group). (F) The representative flow plots represents the percentage of CD11b + Ly6G + neutrophils and CD64 + MHCII + macrophage in the WT and Il1r1 -/- mice skin tissue infected with 0387 pmr1Δ . The bar graph represents the percentage and absolute number of CD11b + Ly6G + neutrophils after (G) 3 DPI and (H) 12 DPI and the CD64 + MHCII + macrophage after (I) 3 DPI and (J) 12 DPI in the WT and Il1r1 -/- mice skin tissue infected with 0387 pmr1Δ ( n = 5–10 mice/group). For C . auris 0387 pmr1Δ infection 5 × 10 6 CFU per mice were used. (K) The bar graph represents the fungal survival of C . auris 0387 and 0387 pmr1Δ primed with neutrophils isolated from bone marrow of WT and Il1r1 -/- mice. ( n = 6) (L) Measurement of IL-1Ra levels from the culture supernatant of BMDM uninfected or infected with live C . auris 0387 or 0387 pmr1Δ for 16 h. ( n = 12). Error bars represent mean ± SEM. * p < 0.05, ** p <0.01, *** p <0.001, **** p <0.0001, NS, non-significant. Abbreviations—DPI, days post-infection; WT, wild type; BMDM, bone marrow-derived macrophages. Statistical significances were calculated using Mann–Whitney U.

Article Snippet: IL-1Ra production in skin homogenate, culture supernatant, and serum was quantified using a mouse IL-1Ra/IL-1F3 DuoSet kit (RnD Systems) by following the manufacturer’s standard operating protocol.

Techniques: Infection, Isolation, Derivative Assay, MANN-WHITNEY