Journal: European Journal of Immunology
Article Title: A Novel Murine Model of Hemophagocytic Lymphohistiocytosis‐Like Inflammation in ZNFX1 Deficiency
doi: 10.1002/eji.70141
Figure Lengend Snippet: BMDMs from Znfx1 mut mice produce more pro‐inflammatory cytokines in an unstimulated state. (A) Schematic representation of the culture and stimulation of bone marrow‐derived macrophages (BMDMs). (B) TNF‐⍺ expression of BMDMs from WT and Znfx1 mut mice in an unstimulated control setting or after stimulation with 2 µg/mL Poly(I:C) HMW for 8 h, with brefeldin A for 6 h. (C) IL‐6 expression of BMDMs from WT and Znfx1 mut mice in an unstimulated control setting or after stimulation with 2 µg/mL Poly(I:C) HMW for 8 h, with brefeldin A for 6 h. (D) IFN‐γ expression of BMDM from WT and Znfx1 mut mice in an unstimulated control setting or after stimulation with 2 µg/mL Poly(I:C) HMW for 8 h, with brefeldin A for 6 h. Representative images are shown in panel (i). The measured log10 single‐cell intensity values are combined into violin plots for each mouse, and the number of measured cells per mouse is stated beneath the respective plot in panel (ii). The values for all the mice in the same group were grouped into violin plots in panel (iii). For the statistical analysis, a separate Gaussian mixture model (GMM) was fitted with the cell intensity values for each mouse. A weighted group mean and the covariance were calculated from the number of observations per mouse. Panel (iv) shows the resulting histogram for unstimulated WT cells with the combination of the weighted GMMs for the entire group (black line) and the three components of the fit (shown in dark blue, light blue, and yellow). The means were compared in a two‐tailed t ‐test; p ‐values are indicated. The data represent the results of one experiment, n = 3.
Article Snippet: To stimulate cytokine production, medium with 2 μg/mL Poly(I:C) HMW (InvivoGen) was added for a total of 8 h. BD GolgiPlug protein transport inhibitor (BD) was added to all wells for a total of 6 h. Cells were fixed with prewarmed paraformaldehyde (Electron Microscopy Sciences, end concentration: 4%) for 30 min at room temperature.
Techniques: Derivative Assay, Expressing, Control, Two Tailed Test