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4 protocols using carboplatin

1

Dual Doxycycline and Carboplatin Murine Protocol

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Doxycycline (Clontech 631311) was administered to mice in water, using 5% sucrose as the vehicle. 2 mg/ml Doxycycline solution was freshly prepared twice a week. Carboplatin (MP Biomedicals 198873) was resuspended in 0.9% NaCl and i.p. injected into mice once a week, at a concentration of 50 mg/kg.
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2

Murine Cholesterol Diet Protocols

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HCD (high cholesterol diet, 2% cholesterol and 0.5% sodium cholate) and CD (control diet, 0.5% sodium cholate) were purchased from TestDiet (5CNY and 5BRW). 27HC and GW273297X were synthesized by Sai Life (Hyderabad, India). GSK 2033 (cat#1221277-90-2), sulfamethoxazole (cat#723-46-6) and trimethoprim (cat#738-70-5) were purchased from Sigma-Aldrich. ICI 182,780 was synthesized by Small Molecule Incorporated (Beijing, China). LDL cholesterol was purchased from Lee Biosolutions (lot#W21129, cat#360–10). 17 β-Estradiol (cat#101656) and carboplatin (lot#M9733, cat#198873) were obtained from MP Biomedicals. Anti-PD-L1 (clone 10F.962, lot#615416D1, cat#BE0115) and IgG2b isotype control (Clone LTF-2, cat#BE0090) were purchased from BioXCell. Flow cytometry antibodies were from BD Pharmingen: CD11b (Cat #557686), Ly6C (Cat #553104), Ly6G (Cat #551461), CD3 (Cat #555275), CD4 (Cat #553729), CD8 (Cat #557682), CD44 (Cat#553134), CD69 (Cat#557392). Recombinant mouse GM-CSF (Cat #P01587) and ID6 (Cat #P08505) were purchased from Novoprotein.
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3

Quantitative Analysis of Chemotherapeutic Agents

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Ifosfamide and cyclophosphamide monohydrate pure standards were purchased from Sigma-Aldrich Co® (St. Louis MO, USA) and MP Biomedicals® (Fountain Pkwy, Solon OH, USA), respectively. Standards of etoposide, carboplatin, vincristine sulphate and methotrexate were all purchased from MP Biomedicals® (Fountain Pkwy, Solon OH, USA), and ondansetron from Toronto Research Chemicals Inc® (Toronto, Canada).
LC-MS grade acetonitrile and methanol were obtained from EMD Millipore Co® (Venezuela), HPLC grade ethyl acetate from Merck® (Darmstat, Germany) and propanol from J.T. Baker® (Xalostoc, Mexico). Ammonium formate was purchased from Sigma Adrich® (St. Louis MO, USA). Whatman 903® cotton filter paper (“Guthrie” cards) were bought from GE Healthcare Ltd.® (Cardiff, UK). Milli-Q grade 1 water (Millipore, Molsheim, France) was used to prepare all the solutions and dilutions.
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4

Dual Doxycycline and Carboplatin Murine Protocol

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Doxycycline (Clontech 631311) was administered to mice in water, using 5% sucrose as the vehicle. 2 mg/ml Doxycycline solution was freshly prepared twice a week. Carboplatin (MP Biomedicals 198873) was resuspended in 0.9% NaCl and i.p. injected into mice once a week, at a concentration of 50 mg/kg.
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