As 252424

After baseline testing (behavior) or at the start of the experiment, animals were anesthetized with isoflurane (5% induction; 1%-2% maintenance). A Hamilton syringe attached to calibrated polyethylene tubing and a 30-gauge needle were used to administer agents subcutaneously to the midplantar hind paw. Vehicle (1% DMSO) or a PI3K isoform-specific antagonist and 100 μL, 2% λ-carrageenan (Wako Pure Chemical Industries, Ltd) contained in the same tubing were injected 2 minutes apart. Class 1 PI3K antagonists used were: compound 15-e (mW 313.4; Enzo Life Sciences, Farmington, NY), alpha; TGX 221, (mW 364.4; Cayman Chemical Company, Ann Arbor, MI), beta; Cal-101 (mW 415.42; Selleck Chemicals, Houston, TX), delta; and AS252424 (mW 305.28; Cayman Chemical Company), gamma. Aliquots of antagonists in DMSO were defrosted and diluted just before use. Vehicle and all antagonist solutions were adjusted to pH of 7.2 to 7.4. Despite different reported IC50s, starting or standard doses of each antagonist were 5 nm administered in 50 μL of vehicle. Doses used were based on recent work.^{13 (link),35 (link)}