Cyclophosphamide (CTX) was purchased from ZhongDa Hospital (Nanjing, CHINA). The drug was dissolved in sterile saline and given intraperitonealy (i.p) following 3 schedules: MTD regimen (150 mg/kg, 3 doses the first week, followed by 2 weeks’ break), metronomic schedule 1 (Met-1, 170 mg/kg every 6 days for 2 cycles) [9 (link)], and metronomic schedule 2 (Met-2, 25 mg/kg every other day) [37 (link)].
RPMI-1640 (Life Technologies, Inc., Grand Island, USA); fetal bovine serum (Hyclone, Australia); Trypsin (Solarbio, Beijing, China), Trypan blue (Invitrogen, Carlsbad, California, USA), Human/Mouse TGF-β1 ELISA kit (eBioscience, San Diego, USA); The PrimeScript RT Master Mix (Perfect Real Time) and SYBR Premix Ex Taq II (Tli RNaseH Plus) kits (TaKaRa, Dalian, China); TRIzol (Invitrogen, Carlsbad, California, USA); Primers for GAPDH, CD4, CD8, FoxP3, α-SMA, FSP-1, TGFβRI, TGFβRII and TGFβRIII (Wei Wo Biotech, Nanjing, China).
RPMI-1640 (Life Technologies, Inc., Grand Island, USA); fetal bovine serum (Hyclone, Australia); Trypsin (Solarbio, Beijing, China), Trypan blue (Invitrogen, Carlsbad, California, USA), Human/Mouse TGF-β1 ELISA kit (eBioscience, San Diego, USA); The PrimeScript RT Master Mix (Perfect Real Time) and SYBR Premix Ex Taq II (Tli RNaseH Plus) kits (TaKaRa, Dalian, China); TRIzol (Invitrogen, Carlsbad, California, USA); Primers for GAPDH, CD4, CD8, FoxP3, α-SMA, FSP-1, TGFβRI, TGFβRII and TGFβRIII (Wei Wo Biotech, Nanjing, China).
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