Smz u zoom 1 10 picospritzer 2 microinjection station

Zebrafish experiments were conducted under the UK Home Office project licence PPL 70/7912 and had been approved by the King’s College Ethical Review Committee. Approximately 500 GFP-tagged PCa cells or AsPC-1 cells were injected into the yolk sac using a Nikon SMZ-U zoom 1:10 Picospritzer II microinjection station. The embryos were checked at 24 h post injection to ensure that GFP signal was restricted to the yolk sac xenograft. 3 days post-injection the percentage of embryos with cancer cell tail invasion was calculated.

All work that was conducted using zebrafish were performed under the UK Home Office project licence PPL 70/7912 and approved by the King's College Ethical Review committee. 2 dpf embryos were submerged in 3.5mM MS222, containing penicillin and streptomycin, and ~500 A-375M2 cells were injected into the embryo yolk sac using a Nikon SMZ-U zoom 1:10 Picospritzer II microinjection station. Injected embryos were placed in E3 media (containing penicillin and streptomycin) and incubated at 28°C for 1 hour to recover, then transferred to 35°C for the remainder of the experiment. 4 hours post-injection, embryos that lacked a clear tumour mass within the yolk sac or that had cells outside of the yolk sac were removed and humanely killed using 15mM MS222. The percentage of embryos with A-375M2 cell tail invasion was calculated 4 days post-injection. The embryos were then humanely killed by the addition of 15mM MS222 for 1 hour.