Anti cd117 apc

Primary SVF from EAT, SAT, BAT was resuspended in PBS with 2% FBS, and incubated with FACS antibodies for 30 min on ice. Cells were initially selected by size on the basis of forward scatter (FSC) and side scatter (SSC), following separated on the basis of cell-surface markers using a flow cytometer MoFlo XDP (Beckman Coulter, Brea, CA). To detect beige progenitors in SVF, primary SVF was separated on the basis of cell-surface markers including anti-PDGFRα (CD140a)-APC (1:50, BioLegend, San Diego, CA) and anti-CD34-FITC (1:100, eBioscience, San Diego, CA). To quantify MCs in adipose tissues, to isolate MCs from SVF, and to remove MCs from SVF, we stained and sorted MCs with anti-CD34-FITC (1:100, eBioscience), anti-CD45-PE (1:500, eBioscience) and MC markers anti-CD117-APC (1:200, eBioscience) and anti-FCεR1-PE-CY7 (1:200, eBioscience). The MC-removed SVF and MCs (1×10⁵) were collected for TPH1 mRNA expressional analysis.

Cellular uptake of antagomirs was detected by FACS analysis. Live cells were treated with either a control or a fluorescent-human labeled antagomir. Mean fluorescence intensity was determined in FlowJo. Cell surface marker staining was performed as previously described [37 (link)] using anti- CD117-APC (eBioscience, San Diego, # 17-1171-82), anti-CD11B-PE (eBioscience, San Diego, #12-0112-85), anti-GR-1-PE-Cy5 (eBioscience, San Diego, #15-5931-81). Data was processed using FlowJo Software Package.