Extended Data Fig. 9: Lack of cell death in Er1Lyz2/− BMDMs and HLA-DR levels in THP-1 cells.
From: DNA damage in macrophages drives immune autoreactivity via nuclear antigen presentation
(A.) Immunofluorescence staining of early endosome antigen 1 (EEA1) in Er1Lyz2/− untreated and dynasore-treated BMDMs. White insets indicate a higher magnification on the right (n = 3). (B.) Single- or two-channel images and higher magnifications of Fig. 5a. Magenta arrowheads point to H1+p62+LAMP-1+ foci. White arrows point to cytoplasmic chromatin fragments. (C.) Flow cytometry analysis of wt and Er1Lyz2/− BMDMs stained with annexin V and propidium iodide (PI) for the detection of cell death. Scatter plots are and the percentages of annexin−PI− cells are shown (n = 4). (D.) LAMP-1 protein levels in equally loaded lysosomal extracts isolated from wt and Er1Lyz2/− BMDMs. Ponceau stain was used for normalization. (n = 4, pval=0.0147) (E.) Quantification of MHC-II in untreated and etoposide-treated THP-1 human monocyte cells. An isotype control histogram is shown in gray (n = 4, pval=0.0006). Asterisk indicates the significance set at p-value: *≤0.05, **≤0.01, ***≤0.01 (two-tailed Student’s t-test). Scale bars: 10μm.
