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5 protocols using oxaliplatin s1224

1

Oral L. lactis administration enhances anti-tumor immunity

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All animal experiments were approved by the Institutional Animal Care and Use Committee of CHA University and KLSbio. The mice used here were maintained and handled according to policies approved by CHA University and KLSbio, respectively. Five-week-old female C57BL/6 mice were purchased from Orient Bio (Gyeonggi, Korea). For the syngeneic tumor mouse model, a total of 2 × 105 MC38 cells were subcutaneously inoculated in C57BL/6 mice. The mice were orally administered the L. lactis strain daily and were intra-peritoneally treated with anti-PD-1 mAb (clone RMP1-14; BioXCell, Lebanon, NH, USA) or 3 mg/kg oxaliplatin (S1224, Selleckchem, Houston, TX, USA) on days 1, 4, 8, 11, and 14. The tumor size was monitored three times per week until the study endpoint. The tumor volume was calculated as the length × width2 × 0.5.
Nude mice were purchased from Orient Bio (Gyeonggi, Korea). Nude mice were orally administered the L. lactis strain for 14 days. Then, a total of 2 × 105 CT26, 1 × 106 4T1, and 1 × 106 HCT116 cells were inoculated subcutaneously into the flank of each mouse. The nude tumor mice were administered the L. lactis daily, and the tumor size was monitored three times a week until the study endpoint. The tumor volume was calculated as the length × width2 × 0.5.
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2

Compound Acquisition for Biological Studies

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Oxaliplatin (S1224) was purchased from Selleck. Guanylic acid disodium salt (GMP, GC35159) was purchased from GlpBio. Mycophenolic acid (MPA, HY-B0421), Mycophenolate mofetil (MMF, HY-B0199) and XAV939 (HY-15147) were obtained from MedChemExpress.
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3

Bifidobacterium Enhances Anti-Tumor Immunity

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All animal experiments were performed with approval from the Institutional Animal Care and Use Committee of CHA University (permission number IACIC180010). Mice used in this study were maintained and handled according to policies approved by CHA University. Five-week-old female C57B6/N mice were purchased from the Orient Bio (Gapyeong, Gyeonggi, Korea). For the tumor mice model, mice were orally administered Bifidobacterium strains for 14 days (−14 day). Then, mice were implanted with 2 × 105 MC38 colon adenocarcinoma cells subcutaneously and treated with 3 mg/kg oxaliplatin (S1224, Selleckchem, Houston, TX, USA) or 2 mg/kg anti-PD-1 mAb (clone RMP1-14, BioXCell, USA) intraperitoneally. Syngeneic tumor mice were treated 6 times with anti-PD-1 mAb or oxaliplatin on days 3, 7, 10, 14, 17, and 21. Tumor size was monitored three times a week until the study endpoint. Tumor volume was calculated as length × width2 × 0.5.
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4

Autophagy and DNA Damage Regulation

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Oxaliplatin (S1224) and CQ (S4330) were purchased from Selleck Chemicals (Houston, TX, USA). The antibodies used for western blot were as follows: anti-c-Myc antibody (sc-40, 1:1500, Santa Cruz, CA, USA), anti-ACTB antibody (sc-10731, 1:2000, Santa Cruz), anti-p62 antibody (66184-1-Ig, 1:1000, Proteintech, IL, USA), anti-LC3 antibody (14600-1-AP, 1:1000, Proteintech), anti-cleaved PARP antibody (#5625, 1:1000, Cell Signaling Technology, MA, USA), anti-cleaved-caspase 3 antibody (#9664, 1:1000, Cell Signaling Technology), anti-ATG10 antibody (DF8366, 1:1000, Affinity, OH, USA), anti-γ-H2AX (ab2893; Abcam, Cambridge, MA, USA), anti-ATG2A antibody (23226-1-AP, 1:1000, Proteintech), anti-ATG2B antibody (251551-1-AP, Proteintech), anti-ATG4C antibody (20382-1-AP, 1:1000, Proteintech).
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5

Immunoprecipitation and Western Blot Analysis

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Primary antibodies were: anti‐FLAG (F1804, Sigma‐Aldrich), anti‐HA (51064‐2‐AP, Proteintech), anti‐PKM2 (D78A4, CST) and anti‐Parkin (66674‐1‐Ig, Proteintech), and normal mouse IgG for immunoprecipitation (12‐371, Sigma‐Aldrich). Blotted PVDF membranes were re‐blotted with anti‐GAPDH antibody (10494‐1‐AP, Proteintech) used as loading controls. Protease Inhibitor Cocktail (C0001) was purchased from TargetMol. PKM2 inhibitor Compound 3K (S8616), 5‐fluorouracil (5‐FU) (S1209) and oxaliplatin (S1224) were purchased from Selleck.
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