TGX-221 

The prostate cancer cell lines DU145 and LNCaP are maintained in RPMI-1640 medium, and PC3 cells are maintained in F-12K medium. LNCaP is a PSMA positive cell line, whereas DU145 and PC3 are PSMA negative. Both are supplemented with 10 % fetal bovine serum. Cells are plated in 96-well flat-bottomed plates at a concentration of 5,000 cells per well in 90 μL of growth medium. After 12 h, TGX-221, BL05, or BL05-HA loaded micelles in PBS are added at concentrations of 0, 0.1, 1, 5, 10, 50 or 100 μM. PBS and 10 μL of trichloroacetic acid (TCA) are added to negative and positive control wells, respectively. After 72 h, 10 μL of 55-μM resazurin blue is added to each well and incubated at 37°C for 4 h. After incubation, the resorufin product is measured with a fluorophotometer using an excitation wavelength of 560 nm and an emission wavelength of 590 nm. The IC₅₀ is determined as the midpoint between positive and negative control groups for each plate using GraphPad Prism 5 software.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Rats are randomLy assigned to drug treatment groups consisting of the vehicle propylene glycol (0.25 mL/kg), LY294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; a reversible non-specific PI3K inhibitor; 10 mg/kg), wortmannin (an irreversible non-specific PI3K inhibitor; 5 mg/kg), IC87114 (2-[(6-aminopurin- 9-yl)methyl]-5-methyl-3-(2-methylphenyl)quinazolin-4-one; a PI3K p110δ antagonist; 2.5 mg/kg) and the selective PI3K p110β antagonist TGX221 (2.5 mg/kg). In the tail bleeding experiments, rats are randomLy assigned to drug treatment groups consisting of LY294002 (10 mg/kg), IC87114 (2.5 mg/kg), wortmannin (5 mg/kg), TGX221 (2.5 or 25 mg/kg), heparin (100 U/kg), aspirin (2× 200 mg/kg p.o.)± heparin (100 U/kg), and aspirin (2× 200 mg/kg p.o.) combined with heparin (100 U/kg) and TGX221 (2.5 mg/kg). All drugs, with the exception of aspirin, are administered as a slow (over ≈45-60 s) i.v. bolus of 0.25 mL/kg into the jugular vein. Aspirin (200 mg/kg suspended in 15% gum arabic in water) is administered twice orally (p.o.)-the first dose is given 24 h before the experiment and the second dose 1 h before the start of the experiment.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

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