Fig. 1: Nampt is upregulated by IFNs and TLR ligands in M1 macrophages and TAMs. | Nature Communications

Fig. 1: Nampt is upregulated by IFNs and TLR ligands in M1 macrophages and TAMs.

From: A Stat1 bound enhancer promotes Nampt expression and function within tumor associated macrophages

Fig. 1

a Table showing top 12 genes from BMM LPS/IFNγ vs. media treated cells and WT vs. IFNγR−/− TAMs that were significantly upregulated by this M1 stimulation or were up significantly in WT TAMs. Log2FC log (base 2) of the expression fold change, FDR false discovery rate. b FPKM of Nampt from sequencing experiments described in a. Flow cytometric sort purification gating strategy provided in Supplementary Fig. 1A. c FPKM of Nos2 and Arg1 from M1 stimulated (LPS/IFNγ) vs. M2 stimulated (IL-4) BMMs. Error bars represent SEM for N = 3, or range for N = 2 independent RNA-seq samples. d qPCR of Nampt in BMMs stimulated for 6 h with the indicated PAMPs. e Northern blot analysis of Nampt from IFNγ stimulated BMMs made from B6 WT mice. Size markers are in kB. f Western blot of NAMPT from BMMs treated with IFNγ for 24 h. Size markers are in kDal. g Expression of Nampt in BMMs treated with Ruxolitinib at the indicated concentrations with and without IFNγ stimulation. h Nampt expression in RAW 264.7 cells in which STAT1 expression was deleted via CRISPR/Cas9 targeting of Stat1 (Stat1 CR) or non-targeting controls (WT EV). i Western blot of NAMPT, STAT1, and ACTIN from RAW 264.7 cell lines described in c. j Nampt, k IL6, and l CXCL10 mRNA expression of cells described in h that were stimulated with the indicated PAMPs for 6 h. WT EV STAT1+ non-targeting controls or STAT1− CR deletions via CRISPR/Cas9 targeting are designated on the x-axes by Stat1+ or −, respectively. Data are representative of at least three independent experiments, except for RNA-seq results in ad, j (two independent experiments). Error bars represent SEM for N = 3 biologically independent samples. p values were determined by a two-tailed unpaired t-test: *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.00005. EV non-targeting CRISPR gRNA control. Uncropped images provided in Source Data file in Supplementary information.

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