mouse igg2a r d pe lgr5 2a2 mouse igg1 origene apc  (OriGene)

Journal: Journal of medicinal chemistry

Article Title: Designing Synthetic, Sulfated Glycosaminoglycan Mimetics That Are Orally Bioavailable and Exhibiting In Vivo Anticancer Activity.

doi: 10.1021/acs.jmedchem.2c01511

Figure Lengend Snippet: Figure 5. In vivo CSC targeting by cholesterol-modified G2.2. (A) Schematic of the study. 1 × 105 CD133+/CXCR4+ (Dual Hi) HT-29 cells were injected subcutaneously to generate xenografts in NCr nude mice. Once the palpable tumors were observed, the animals were treated with vehicle (Veh) or FUOX (25 mg/kg 5-fluorouracil (FU) & 2 mg/kg oxaliplatin (OX); IP weekly for 3 weeks) followed by randomization to vehicle (Veh), FUOX (25 mg/kg FU & 2 mg/kg OX IP weekly), or G5C (100 mg/kg three times/week × 4.6 weeks (14 doses)). Animals were sacrificed at the end of treatment and tumors (n = 2 per group) were analyzed for CSCs. (B) Profile of change in tumor volume as the function of the second treatment (Veh, FUOX, or G5C). (C) Animal weight measured in grams at least three times a week. Although weight was measured since initiation of tumors, data is shown only following randomization to NSGM or FUOX. (D) Profile of ex vivo growth of 1, 2, and 3° spheroids from xenograft cells. (E) Quantitation of Dual Hi (CD133+/CXCR4+) cells in xenografts by flow cytometry. (F) Profile of LGR5, DCAMLK1 (CSC markers), and BMI-1 (self-renewal factor) proteins in lysate from xenografts using Western analysis. *p < 0.01 compared to vehicle controls; ¶ p < 0.01 compared to FUOX. Error bars represent 1 ± standard error of the mean (SEM).

Article Snippet: We used the CXCR4 (anti-mouse CD184)-PE-conjugated clone 2B11 antibody (Cat # 12-9991-82, eBioscience, San Diego, CA; dilution 1:50) and the anti-CD133 mouse mAb (Cat # 130-090-826, Miltenyi Biotec, Auburn, CA; dilution 1:33) for quantifying CXCR4+ and CD133+ cells as well as anti-LGR5 mouse-PE antibody (Cat # TA400001, Origene, Rockville, MA; dilution 1:50).

Techniques: In Vivo, Modification, Injection, Ex Vivo, Quantitation Assay, Flow Cytometry, Western Blot

Journal: Journal of medicinal chemistry

Article Title: Designing Synthetic, Sulfated Glycosaminoglycan Mimetics That Are Orally Bioavailable and Exhibiting In Vivo Anticancer Activity.

doi: 10.1021/acs.jmedchem.2c01511

Figure Lengend Snippet: Figure 7. In vivo CSC targeting by orally administered cholesterol analogues of NSGMs. (A) Schematic of the study. 1 × 105 CD133+/CXCR4+ (Dual Hi) HT-29 cells were injected subcutaneously to generate xenografts. Once the palpable tumors were observed, the animals were treated with FUOX (50 mg/kg FU + 5 mg/kg OX; IP weekly for 3 weeks) followed by randomization to either vehicle (Veh) or G2C (100 mg/kg PO; 5 times/week for 3 weeks) [Experiment 1], or vehicle, G2.2 (100 mg/kg IP; 3 times/week for 3 weeks) or G2C (100 mg/kg PO; 5 times/week for 3 weeks) [Experiment 2]. (B, C) Ex vivo analyses of tumor xenograft for anti-CSC vs anti-NSC (normal stem cell) properties. (B) Profile of ex vivo growth of 1, 2, and 3° spheroids from xenograft cells. (C) Profile of ex vivo growth of intestinal crypt-derived organoids cultured in Matrigel in the presence of IntestiCult media. (D) Relative anti-CSC efficacy of parental G2.2 and oral G2C. (E) Growth curves showing tumor volume inhibition by oral G2C and parental G2.2. Quantitation of Dual Hi (CD133+/CXCR4+) and LGR5+ cells (CSCs) in xenografts by flow cytometry. # ptrend < 0.1, * p < 0.05, ** p < 0.01 compared to vehicle control. Error bars represent ±1 SEM.

Article Snippet: We used the CXCR4 (anti-mouse CD184)-PE-conjugated clone 2B11 antibody (Cat # 12-9991-82, eBioscience, San Diego, CA; dilution 1:50) and the anti-CD133 mouse mAb (Cat # 130-090-826, Miltenyi Biotec, Auburn, CA; dilution 1:33) for quantifying CXCR4+ and CD133+ cells as well as anti-LGR5 mouse-PE antibody (Cat # TA400001, Origene, Rockville, MA; dilution 1:50).

Techniques: In Vivo, Analogues, Injection, Ex Vivo, Derivative Assay, Cell Culture, Inhibition, Quantitation Assay, Flow Cytometry, Control