Gene expression is imaged in deep tissues in mice using ultrasound
-AxeTxe in EcN, the glycerol stock of this strain was used to inoculate 2 ml of LB supplemented with 25 μg mlof chloramphenicol and 1% glucose, and this culture was incubated at 37 °C and 250 r.p.m. overnight. Then, 20 μl of the overnight culture was subcultured into 2 ml of LB with 25 μg mlof chloramphenicol, 2 ml of LB without antibiotics and 2 ml of LB without antibiotics and with 0.1% L-arabinose, each in quadruplicate.
of 1–2, or liquid cultures were used directly. Suspensions were transferred to glass slides, and PCM images were acquired using a Zeiss Axiocam microscope with a ×40 Ph2 objective.All in vivo experiments were performed under protocol 1735 or 1761, approved by the Institutional Animal Care and Use of Committee of the California Institute of Technology. Animals were housed in a facility maintained at 71–75 °F and 30–70% humidity, with a lighting cycle of 13 hours on and 11 hours off .
of 0.625 and used for injection. Three days after injection of EcN, mice were injected intraperitoneally with 120 mg of L-arabinose to induce the EcN. Starting 24 hours after induction, ultrasound images of tumors were acquired as described in Supplementary Note. After imaging, 3.0-MPa acoustic pressure was applied throughout the tumor to collapse GVs, and mice were injected again intraperitoneally with 120 mg of L-arabinose.
The lentiviral transfer plasmid with constitutively expressed tetracycline transactivator was constructed as follows: pNCS-Antares was obtained from Addgene , and P2A was added to the N-terminus of Antares with a primer overhang during PCR. This fragment was subcloned into the lentiviral transfer plasmid pEF1α-rtTA-WPRE between rtTA and WPRE in-frame with the rtTA ORF using NEBuilder HiFi DNA Assembly.
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