4T1 mouse breast carcinoma cells were suspended in RPMI medium/Matrigel (1:1) (1 × 106 in 50 μL) and injected into the mammary pad of female BALB/c mice under depth anesthesia, as previously described [15 (link)]. At two weeks post tumor cells inoculation, mice had been started to be treated via intraperitoneal injection (twice per week) with 10% (100 µg/mL) Mn-CDs-NHF (n = 6) or Gadovist (n = 6) for 3 weeks. CD-NHF concentration is relative to mice blood volume and represents one of the previously tested concentrations [15 (link)].
At the end of the testing period, the animals were euthanized (neck dislocation under deep anesthesia), and primary tumors and various organs (liver, kidneys, lungs, spleen) were collected. Half of each primary tumor and half of each lung were immediately suspended in RNA Save (Biological Industries, New Haven, CT, USA) and stored at −80 °C until use. The other half from each primary tumor and each lung were preserved in 10% paraformaldehyde (Sigma-Aldrich) for further analysis.
At the end of the testing period, the animals were euthanized (neck dislocation under deep anesthesia), and primary tumors and various organs (liver, kidneys, lungs, spleen) were collected. Half of each primary tumor and half of each lung were immediately suspended in RNA Save (Biological Industries, New Haven, CT, USA) and stored at −80 °C until use. The other half from each primary tumor and each lung were preserved in 10% paraformaldehyde (Sigma-Aldrich) for further analysis.
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