Rat anti mouse f4 80 fitc

BM cells were flushed from femur and tibia. An aliquot of 1 × 10⁶ cells was stained with the following fluorescence-conjugated antibodies: Rat anti-Mouse F4/80 FITC, Rat IgG2aκ (isotype control), Rat anti-Mouse CD11b PE, Rat IgG2bκ (isotype control), all from eBiosciences, San Diego, CA, USA. Cells were incubated with the antibodies for 30 min at 4 °C and washed with phosphate buffered saline (PBS) containing 2% fetal bovine serum (FBS). Cells were fixed with 1% paraformaldehyde (PFA)+PBS and analyzed on a FACSort flow cytometer (Becton-Dickinson, New Jersey, USA). Results were analyzed using Kaluza software (Beckman Coulter, Nyon Switzerland).

16:4(n-3) was isolated from Ulva pertusa as previously described (23 (link)). GW1100 was purchased from Cayman Chemical (Ann Arbor, MI, USA). GW9508 and AACOCF₃ (arachidonyl trifluoromethyl ketone) were purchased from Tocris (Bristol, United Kingdom). NCG21, TUG-891, AH-7614, and TUG-1197 were synthesized as described previously (24 (link)–27 (link, link, link)). For fluorescence-activated cell sorting (FACS) analysis, the following Abs were used: rat anti-mouse F4/80-FITC and rat anti-mouse CD11b-APC (both from eBioscience, San Diego, CA, USA). For immunohistochemical staining, the following Abs were used: anti-γH2AX (gamma histone 2A family member X2577; Cell Signaling Technologies, Danvers, MA, USA), anti-GPR120 (NBP1-00858; Novus Biologicals, Littleton, CO, USA) and poly–horseradish peroxidase (HRP) goat anti-rabbit/rat/mouse (Immunologic, Duiven, The Netherlands). For Western blotting, the following Abs were used: rabbit anti–phospho-cPLA₂ (Ser505, 2831; Cell Signaling Technologies), mouse anti–β-actin (NB600-501; Novus Biologicals), and goat anti-mouse HRP (Santa Cruz Biotechnology, Santa Cruz, CA, USA).