Fig. 5: Protein content alterations in the antigen presentation compartment of Er1^Lyz2/− macrophages.

a,b, Co-localization studies of autophagy (p62), lysosomes (LAMP-1) and histone H1 (a) or lamin B1 (LMNB1) (b) upon chloroquine or chloroquine and Dynasore (Dyn) treatment. Magenta arrowheads point to H1⁺p62⁺LAMP-1⁺ (a) or LMNB1⁺p62⁺LAMP-1⁺ (b) foci. White arrows point to cytoplasmic chromatin fragments. Single-channel or two-channel images and higher magnifications of a are shown in Extended Data Fig. 9b and Supplementary Fig. 2a,b. The two graphs illustrate the percentage of cells with any triple co-localized foci (n = 3 biological replicates, 4−7 optical fields) or the total number of triple co-localized foci per each individual cell counted (n ≥ 144 cells for H1 and n ≥ 162 cells for LMNB1). c, Western blot analysis of lysosomes purified from WT and Er1^Lyz2/− BMDMs, after chloroquine treatment for 3 h. Membranes were probed for ribosomal (FAU) and nuclear (H1, LMNB1 and LMNA/C) markers. LAMP-1 was used as a resident protein of lysosomes. The splice point is indicated by a vertical black line. Equal amounts of protein (up to 10 μg) were loaded, and protein levels were normalized according to Ponceau stain. The normalized (Norm.) protein levels are plotted (n = 3−4). d,e, Immunofluorescence staining of THP-1 human monocytes (n = 3, P = 0.0053) (d) and cells of monocytic origin sorted from NZB/NZW F₁ mice (SLE) (n = 3 P = 0.0115) (e), stained for DAPI, H1 and p62. Cells were fixed after a 3-h treatment with chloroquine. Magenta arrowheads point to chromatin fragments co-localized with p62. The graphs illustrate the percentage of cells with cytoplasmic chromatin fragments. Error bars indicate s.e.m. among replicates. *P ≤ 0.05, **P ≤ 0.01 and ****P < 0.0001 (two-tailed Student’s t-test). Scale bars, 10 μm. Exact P values are provided in the Source Data.

Source data