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. 2018 Oct 22;9(11):1081. doi: 10.1038/s41419-018-1125-5

Fig. 6. Macrophage-derived TNFα and in vivo efficacy of TL32711.

Fig. 6

a Human TNFα ELISA quantifying TNFα secretion from THP1 cells (THP basal) and M1-differentiated THP1 cells(THP M1) at 24 h. b Annexin-V/PI flow cytometric analysis of PC3 cells pretreated for 24 h with 2.5 μM Entinostat(Ent) followed by 24 h with 1 µM TL32711 cultured in M1-derived conditioned mediain the presence(CM + TNFα nAb) or absence(Cond Media) of 100 ng/mL TNFα neutralising antibody. c Tumor volume analysis of PC3 xenografts implanted into SCID-mice treated with Vehicle(Veh), 20 mg/kg TL32711, 15 mg/kg Entinostat(ENT) or TL32711 and Entinostat combination(schedule as indicated; E- Entinostat, T- TL32711). d Mouse weights for duration of xenograft study. *p ≤ 0.05, **p ≤ 0.01 and ***p ≤ 0.001. e Western blot analysis of cIAP1 expression in PC3 cells pre-implantation (in vitro) and three individual PC3 xenograft tumor lysate 24 h following final dose of Vehicle (Veh), Entinostat(Ent), TL32711 or Combination(Combo). f Immunohistochemical analysis of FLIP (Left), and cleaved caspase-3 (Right) expression in PC3 xenograft tumors from mice 24 h following final dose of Vehicle, Entinostat, TL32711 or Combination. Representative IHC images of 3 regions of 3 tumors collected per treatment group. g Immunoflurorescent microscopy of the murine macrophage marker F4/80, cytokeratin (CK)-5 epithelial cell marker, and DAPI-stained nuclei in PC3 xenograft tissue. Scale bar represents 100 μm. h Murine TNFα ELISA quantifying murine TNFα present in the serum of mice 24 h following final dose of Vehicle (Veh), Entinostat (Ent), TL32711 or Combination(Combo). Data represents pooled values in duplicate from 3 mice per treatment group. *p ≤ 0.05, **p ≤ 0.01 and ***p ≤ 0.001