Figure 3.

Induction of NF-κB activity in reporter mice after administration of M-VM3. (a) M-VM3 induces long-term activation of NF-κB in live mouse hepatocytes carrying an introduced NF-κB-dependent luciferase reporter construct. Cells were infected with M-VM3 (MOI=10⁴) or Ad-mCherry (MOI=10⁴) or treated with entolimod (0.1 mg/ml) or PBS (control), then these agents were removed from the media (3 h for Ad and 1 h for entolimod) and luciferase was measured by LumiCycle. The level of luciferase activity from PBS-treated cells was subtracted. (b) BALB/C-Tg(IkBa-luc)-Xen mice were given a single intraprostate injection of PBS, CBLB502 (1 μg per mouse) or M-VM3 (1 × 10⁹ v.p.) and analyzed 3, 24 or 48 h later by whole-body Xenogen bio-luminescence imaging of live anesthetized animals. (c) Measurement of luciferase activity in liver (L), intestine (I) and prostate tissue (P) extracts of NF-κB-luciferase reporter mice BALB/C-Tg(IkBa-luc)-Xen after intravenous and intraprostate injections (48 h) of M-VM3. Relative light unit (RLU) values (per mg of total protein) in tissue extracts of M-VM3-treated mice were calculated by subtraction of RLU values for PBS-treated mice.