Anti cd130 bv421

The viability of disaggregated primed and naive hESCs and hiPSCs was analyzed with a Life/Death Fixable kit (Invitrogen, Waltham, MA, USA) in a FACS Canto II (BD Biosciences, Franklin Lakes, NJ, USA). Cells were stained with anti-CD24 PE (BD Biosciences, clone ML5) and anti-CD57 APC (BD Bioscience, clone NK-1) primed markers with anti-CD75 FITC (BD Biosciences, clone LN1) and anti-CD130 BV421 (BD Biosciences, clone AM64) naive markers and anti-CD90 PE-Cy7 (BD Biosciences, clone 5E10) stemness marker. Approximately 30,000–50,000 events were acquired for analysis. Populations were analyzed using FlowJo v.X.0.7 (TreeStar Inc., Ashland, OR, USA). See Supplementary Table S1.

ASC were immunophenotypically characterized by staining for CD45-FITC, CD31-FITC, CD13-PECy7, CD73-PE, and CD90-APC (all BD Biosciences, San Jose, CA). For detection of IL-6 and IFN-γ receptors, 400,000 ASC (n = 3) were stained for two IL-6 receptor subunits (CD126 and CD130) and IFN-γ receptor (CD119). The cells were incubated with anti-CD126-PECy7 (BioLegend, San Diego, CA), anti-CD130-BV421 (BD Biosciences, San Jose, CA), anti-CD119-APC (SB Sino Biological Inc., Beijing, China), or isotype-matched control antibodies (eBioscience, San Diego, CA) in the dark for 30 min at room temperature. Thereafter, the cells were washed twice with FACSFlow (BD Biosciences) and measured on a FACS Canto II flow cytometer (BD Biosciences) and analyzed with Kaluza Analysis 1.3 software (Beckman-Coulter, Brea, CA).