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. Author manuscript; available in PMC: 2019 Aug 18.
Published in final edited form as: Nat Cell Biol. 2019 Feb 18;21(3):397–407. doi: 10.1038/s41556-019-0287-4

Figure 4. NMN enhances the inflammatory environment and cancer progression in vivo.

Figure 4.

a, In established senescent cells, HMGA1 or NAMPT were knocked down or cells were treated with FK866 to assess the effect on the growth of co-cultured luciferase-expressing TOV21G ovarian cancer cells. Cell growth was assessed by luminescence following eight days of growth. (n=3 independent experiments). b-n, Wildtype mice were compared to KC mice treated with vehicle, NMN (500 mg/kg daily), or FK866 (25 mg/kg daily). Representative H&E images of pancreas (b) and quantification of percent acinar area (c). Representative Masson trichrome staining images of pancreas (d) and quantification of percent trichrome area (e). Expression of IL1β, IL-6 and IL-8 was determined using qRT-PCR analysis (f). Representative immunohistochemical staining of infiltrating F4/80-positive immune cells (g) and quantification of percent F4/80 positive cells (h). Representative immunohistochemical staining of infiltrating CD3-positive immune cells (i) and quantification of the number of CD3 positive cells/field (j). Representative SA-β-gal staining (k) and quantification of SA-β-gal positive areas (l) in the indicated treatment groups. Expression of p16 (m) and p21 (n) was determined using qRT-PCR analysis. n=10 mice/group unless otherwise stated. Scale bar for all images is 200 μm. o, Immunoblot of the indicated protein in TOV21G cells containing doxycycline-inducible knockdown of NAMPT with or without doxycycline treatment. p, TOV21G and oncogene-induced senescent IMR90 cells were subcutaneously co-injected into the right dorsal flank of 6-8 week old NSG female mice. The mice (n=9 mice/group) were treated with vehicle control, NAM (500 mg/kg; intraperitoneal injection; every other days) for 17 days. Tumor growth in the indicated treatment groups was measured at the indicated time points. All graphs represent mean ± s.d. P values were calculated using a two-tailed t-test. Statistical source data are provided in Supplementary Table 1. Unprocessed original scans of all blots with size marker are shown in Supplementary Fig. 8.