Additional file 1 of Role of PD-L1 in licensing immunoregulatory function of dental pulp mesenchymal stem cells

Additional file 1: Figure S1. Gating strategy to determine PD-1 and PD-L1 expression by DPSCs. DPSCs were identified by FSC and SSC (1) then live cells were gated by SSC/dead staining dot plot (2). Afterwards CD3neg CD4neg cells were selected (3) and fluorescence intensities in FITC and PECy7 channels shown by histograms. The mean percentage of adherent DPSCs in the FSC/SSC gating was 93.5% �� 2.2%. Figure S2. Gating strategy to determine PD-1 and PD-L1 expression by PBMCs pre-activated by CD3/CD28 linking. PBMCs were identified by FSC and SSC (1) then live cells were gated by SSC/dead staining dot plot (2). Afterwards CD3+CD56neg cells (T lymphocytes) were selected (3). CD4+ and CD4neg cells were further gated (4) and fluorescence intensities in FITC and PECy7 channels shown by histograms. Figure S3. Real Time PCR analysis of caspase 3 in PBMCs. Caspase 3 mRNA levels were evaluated in rPBMCs and aPBMCs alone, aPBMCs after DPSCs co-culture with and without PD-L1 inhibitor. Statistically significant increase of mRNA levels of caspase 3 was detected in aPBMCs after DPSCs co-culture with and without the addition of PD-L1 inhibitor (��P < 0.05 vs aPBMCs).