Fig. 1: DNA damage accumulation in Er1F/− macrophages.
From: Tissue-infiltrating macrophages mediate an exosome-based metabolic reprogramming upon DNA damage
a Lys-Cre-driven Rosa-YFP expression in thioglycolate-elicited peritoneal macrophages (TEMs; n > 500 cells counted per genotype). The numbers indicate the average percentage of GFP (+) cells ± SEM), b Lys-Cre-driven Rosa-YFP expression in hepatocytes and primary pancreatic cells (PPCs) shown by confocal microscopy (n > 100 cells counted per genotype; the numbers indicate the average percentage of GFP (+) cells ± SEM) and c western blotting. d Immunofluorescence staining of Lys-Cre-driven Rosa-YFP expression in the Er1F/− pancreas and the white adipose tissue (WAT) that are infiltrated with MAC1-positive macrophages (indicated by the arrowheads). e Western blotting of ERCC1 protein in whole-cell (w) cytoplasmic (c) and nuclear (n) extracts. Tubulin (TUB), and Fibrillarin (FIB) were used as loading controls (as indicated). The graph represents the fold change (F.C.) of ERCC1 protein levels in Er1F/− samples compared to corresponding Er1F/+ controls (n = 3). f Cell type-specific ablation of ERCC1 (indicated by the arrowhead) in bone marrow-derived (BMDMs) and TEMs expressing the macrophage-specific antigen MAC1. The numbers indicate the average percentage of ERCC1 (+) nuclei ± SEM in Er1F/+and Er1F/− BMDMs and TEMs (n > 150 cells were counted per genotype). g Immunofluorescence detection of γ-H2AX in Er1F/− and Er1F/+ BMDMs and h TEMs. i Immunofluorescence detection of FANCI, pATM and RAD51 in Er1F/− and Er1F/+ BMDMs (in each case n > 200 cells were counted per genotype). j Immunofluorescence detection of pATM in Er1F/− and Er1F+ TEMs (n > 150 cells were counted per genotype). k Immunofluorescence detection of Caspase 3 (CASP3) (n > 300 cells were counted per genotype) and l GL13 (indicated by the arrowhead), commercially available SenTraGor®, in Er1F/− and Er1F+ BMDMs. Fluorescence intensity was calculated in n > 50 cells per genotype. Gray line is set at 5 μm scale, unless otherwise indicated. Error bars indicate S.E.M. among replicates (n ≥ 3). Asterisk indicates the significance set at p-value: *≤0.05, **≤0.01 (two-tailed Student’s t-test).
