Journal: Science Advances

Article Title: Mannan-induced Nos2 in macrophages enhances IL-17–driven psoriatic arthritis by innate lymphocytes

doi: 10.1126/sciadv.aas9864

Figure Lengend Snippet: ( A ) The numbers of CD11b + CD45 + live skin cells were calculated for hind paws from both Ncf1 +/+ and Ncf1 */* mice with l -NAME or PBS treatment at 5 DPI. n = 6 mice per group including the naïve mice. ( B to D ) The concentrations of TNF-α, IL-17A, and IL-1α in the supernatant solution of skin cells were quantified after 4 hours of stimulation with PMA/ionomycin. ( E ) The NOS2-dependent increase of IL-1α was found in the supernatant of skin cells from the mice with MIP disease at day 5, and each group included five mice for the cytokine analysis. ( F ) Evaluation of the time course was performed in Ncf1 */* mice during IL-1α in vivo neutralization ( n = 5 mice per group) after mannan injection. All results are shown as mean ± SEM.

Article Snippet: Cytokine levels in the skin culture supernatant were measured by flow cytometry using BD Cytometric Bead Array Mouse/Rat Soluble protein Master Buffer kit (IL-1α, TNF-α, and IL-17A) according to the manufacturer’s instruction.

Techniques: In Vivo, Neutralization, Injection

Journal: Science Advances

Article Title: Mannan-induced Nos2 in macrophages enhances IL-17–driven psoriatic arthritis by innate lymphocytes

doi: 10.1126/sciadv.aas9864

Figure Lengend Snippet: The Ncf1 */* mice received an arthritogenic dose of mannan at day 0. l -NAME and PBS were injected intraperitoneally daily for 5 days beginning on day 0, and the local skin was excised from hind paws when the mouse was sacrificed using CO 2 at day 5. The skin cells were analyzed by flow cytometry for the following cell populations: (i) total number of CD45 + live cells, (ii) CD45 + CD3 − CD90.2 + CD127 + RORγt + live cells (ILC3), and (iii) CD45 + βTCR − γδTCR + live cells (γδ T cells). ( A ) Schematic illustration of the region of interest (ROI) at the position corresponding to dotted lines for isolating skin samples from the paw. ( B ) Number of CD45 + live skin cells from the selected ROI in the paw. ( C to F ) Frequency of IL-17A + expression was shown within both ILC3 in (C) and (D) and γδ T cells in (E) and (F). Each group included five mice for the intracellular cytokine analysis by flow cytometry. All results are shown as mean ± SEM. Cy7, Cyanine7.

Article Snippet: Cytokine levels in the skin culture supernatant were measured by flow cytometry using BD Cytometric Bead Array Mouse/Rat Soluble protein Master Buffer kit (IL-1α, TNF-α, and IL-17A) according to the manufacturer’s instruction.

Techniques: Injection, Flow Cytometry, Expressing