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. 2019 Dec 4;5(12):eaaw9051. doi: 10.1126/sciadv.aaw9051

Fig. 6. TNFα is required for prepolarization of DC1 from BM throughout C. neo infection.

Fig. 6

BM was harvested from control or αTNFα mice at the indicated times after infection. (A) Magnetically separated BM DC precursors were stained for MLL1 (green), H3K4me3 (red), and 4′,6-diamidino-2-phenylindole (DAPI) (blue). Representative images were taken at 40×. Fluorescence intensity was measured by imageJ and normalized to DAPI between images. n = 6 independent wells from separate matched experiments; three random fields were analyzed from each independent well. (B) FACS (fluorescence-activated cell sorting)–sorted DC precursors were stained for intranuclear H3K4me3; representative histograms of 14 dpi with cumulative bar graph of MFI throughout infection are shown. n = 8 from separate matched experiments. ***P < 0.001. (C and D) BMDCs were matured for 7 days ex vivo with GM-CSF from BM of uninfected control and αTNFα mice (C) or 7 dpi control and αTNFα mice (D) and then treated as in Fig. 1. DC1 gene stability was assessed. Data are normalized to control DC1 values within each graph. n = 6 from separate matched experiments. *P < 0.05 by ANOVA within uninfected control and uninfected αTNFα and ‡P < 0.05 by ANOVA between infected control and αTNFα. n.s., not significant.