Rediject d luciferin

All obtained mice were acclimatised for at least 1 week. Vendor health reports indicated that the mice were free of known viral, bacterial and parasitic pathogens. 5 × 10⁴ R211‐Luc cells were injected into the tail vein of female 8‐week‐old nude mice (Charles River). The mice were treated 5 days a week with vehicle or BYL‐719 administered by oral gavage at 50 mg/kg daily for 3 weeks starting on the injection date. Two‐ and three‐week post‐injection, mice were injected i.p. with 150 mg/kg of RediJect D‐Luciferin (Perkin Elmer) and monitored for Luciferase expression after 6–8 min in the IVIS Spectrum in vivo imaging system (Perkin Elmer). Luminescence (photo count) was measured for each mouse. Lungs were inflated then fixed and embedded in paraffin. 1 × 10⁵ A338 or 1 × 10⁵ A260 cells were injected into the tail vein of female 8‐week‐old C57/B6 mice (Charles River). Three‐week post‐injection, lungs were inflated then fixed and embedded in paraffin.

Male 12-week-old NSG mice (NOD.Cg-Prkdcsid Il2rgtm1Wjl/SzJ) were purchased from the Jackson Laboratory (Bar Harbor, ME) and used under a Stony Brook University IACUC-approved protocol. NSG mice were irradiated with a sublethal (2.0 Gy) dose of gamma irradiation. Next day, mice were intravenously (IV) injected with 1.0 x10⁶ Jurkat cells that had been stably transduced to express luciferase, in order to cause a measurable IV tumor to form. Three days (Day 3) following Jurkat cell injection, mice were intravenously injected via tail vein with one course consisting of 15 × 10⁶ CD3CAR NK-92 cells or vector control NK-92 cells (N = 6 per group) during the window of the NK cell life expectancy and concluding by Day 10. Upon injection of NK-92 cells on Day 10, 2 mice subsequently died within 30 minutes after injection procedure. Two additional low dose injections totaling 5 x10⁶ CD3CAR NK-92 cells was administered through Day 14 and 23 to see if this tumor control could be maintained. On days 4, 7, 9, and 13, mice were injected intraperitoneally (IP) with 100 μL RediJect D-Luciferin (Perkin Elmer, Waltham, MA) and subjected to IVIS imaging (PerkinElmer, Waltham, MA). Images were analyzed using Caliper Life Sciences software (PerkinElmer, Waltham, MA).

Bioluminescence was detected at different time-points including 5 h, 1 day, 2 days, 3 days, 6 days and 8 days after Luc-mRNA-LNP injection. Mice were anesthetized in a chamber with 2.5% isoflurane and given intra-peritoneal injections of RediJect D-Luciferin (Perkin Elmer, Waltham, MA, 150 mg/kg). Luminescence was detected with an IVIS Spectrum imaging system (PerkinElmer, Waltham, MA) while maintaining 2% isoflurane in the imaging chamber via a nose cone. Images were captured 10 min after luciferin administration with sequence set-up of 1 s, 10 s and 20 s. The photon flux values (photons/second), corresponding to the region of interest marked around the bioluminescence signal, were analyzed using the Caliper Life Sciences software (Living IMAGE Software, Caliper).