Female balb c nu nu mice

Male BALB/c mice and female BALB/c-nu/nu mice were purchased from Orient Bio, Inc. (Sungnam, Korea) and maintained under temperature-, humidity-, and light-controlled conditions. All animal experimental protocols were approved by the institutional review board and performed in accordance with the NIH Guide for the Care and Use of Laboratory Animals.²² Mouse hepatoma Hepa-1c1c7 cells were obtained from the ATCC (Rockville, MD) and cultured in Dulbecco's modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum. The fusion protein R-III (Supplemental Figure S1) was synthesized using a previously described method.¹⁹ (link)

Female BALB/c nu/nu mice (5–6 weeks old) were obtained from Orient Bio Inc. (Seongnam‐si, Gyeonggi‐do, Korea). Animal care and experimental protocol was in accordance with the guidelines of the Institutional Animal Care and Use Committee (IACUC) of College of Medicine, The Catholic University of Korea (CUMC‐2010‐0141‐01). For tumor induction, 5 × 10⁶ viable Capan‐1 cells were injected s.c. into the right flank of mice. Drug treatment was started when tumor size reached approximately 200 mm³. For the combination study, PTX and GEM were given i.p. at 100 mg/kg per day and 50 mg/kg twice a week, respectively, for 4 weeks. For the combination of PTX and DOX (or LP‐DOX), PTX was given i.p. at 100 mg/kg per day for 2 weeks prior to i.v. administration of DOX and LP‐DOX at 8 mg/kg by tail vein once. For the drug distribution experiment, a higher dose of 20 mg/kg as DOX was given after PTX pretreatment, and tumor tissue was harvested after 5 min (DOX) or 24 h (LP‐DOX) when the maximum drug distribution was expected in each group. Excised tumors were divided into several pieces and were either fixed overnight in 10% formalin solution for immunohistochemistry or embedded in OCT compound (Sakura, Tissue‐Tek^®, Torrance, CA, USA) for immunofluorescence staining. For biochemical analysis, some tumor pieces were snap‐frozen and stored at −70°C until analysis.

Female Balb/c (nu/nu) mice, 5 weeks of age, were purchased from Orientbio (Seoul, Korea). All animal experiments were conducted in accordance with the Guide for the Care and Use of Laboratory Animals, and approved by the Institutional Animal Care and Use Committee of Seoul National University. After 1 week of adaptation, MDA-MB-231 cells with control or TGF-β-activated CCD-1068sk fibroblasts were co-injected subcutaneously into the flank of mice as described previously [11 (link)].