Taqman openarray mouse phagocytosis panel

Reagents for reverse transcription (High-Capacity cDNA Reverse Transcription Kit with RNase Inhibitor) and quantitative PCR (Taqman Assays, TaqMan OpenArray Mouse Phagocytosis Panel, and TaqMan Fast Advanced Master Mix), red fluorescent microspheres (FMS), 2.0 μm, Hoechst 33342, CellMask Orange Actin Tracking Stain, CellMask Green Actin Tracking Stain, RPMI were obtained from Thermo Fisher Scientific, Inc. (Waltham, MA, USA). Opti-MEM was from Gibco, negative siRNA control was from Ambion, Lipofectamine was from Thermo Fisher Scientific, Inc., Aβ_1–42 and Aβ_1–42 HiLyte 488 were from AnaSpec, Inc. (Fremont, CA, USA). JQ1, GSK12101517, IBET-762, OTX-015, PFI-1, Cytochalasin D were obtained from Sigma-Aldrich (St. Louis, MO, USA). Heat-inactivated foetal bovine serum (FBS), Accutase solution, penicillin, streptomycin, L-glutamine, 3-(4,5-dimethyl-2-tiazolilo)-2,5-diphenyl-2H-tetrazolium bromide (MTT), propidium iodide, TRI-reagent, DNase I, dithiothreitol (DTT), anhydrous dimethyl sulfoxide (DMSO), lipopolysaccharide from Escherichia coli O55:B5 (toxicity 3,000,000 U/mg), and all other reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA).

The level of mRNA for phagocytosis-related genes was analysed using TaqMan OpenArray Mouse Phagocytosis Panel (Catalog number: 4471126) according to the manufacturer’s protocol (Thermo Fisher Scientific, Inc.). An amount of 40 ng of cDNA was used per each well. Data were analysed and calculated with Expression Software version 1.3 (Thermo Fisher Scientific, Inc.) using global normalization and the relative levels of mRNA were calculated using the ΔΔCt method.