Raw data for Figure 4A: Neutralising antibodies against IFN-γ and TNF-α reduce the immunosuppressive capacity of Mesoangioblasts/HIDEMs

CFSE labelled PBMCs were stimulated with anti-CD3/CD28 beads in the presence of HIDEMs/mesoangioblasts (1:4) and neutralising antibodies against IFN-γ and TNF-α or irrelevant isotype control antibody (0.5, 1.0 and 2.0 µg/ml) or recombinant IL-1RA (0.5, 1.0 and 2.0 µg/ml). Cells were harvested on day 6 and stained with anti-CD3 and 7AAD. After gating on CD3+7AAD- the number of CFSE diluting cells were enumerated using counting beads. Experiments were carried out in duplicates. n=4.

将经CFSE（羧基荧光素琥珀酰亚胺酯）标记的外周血单个核细胞（Peripheral Blood Mononuclear Cells，PBMCs）与HIDEMs、成血管间充质细胞（mesoangioblasts）按1:4的比例混合，以抗CD3/CD28磁珠进行刺激，同时分别添加针对干扰素-γ（IFN-γ）与肿瘤坏死因子-α（TNF-α）的中和抗体、同型无关对照抗体（浓度设为0.5、1.0及2.0 μg/ml），或重组白细胞介素1受体拮抗剂（recombinant IL-1RA，浓度同样为0.5、1.0及2.0 μg/ml）。于培养第6天收集细胞，采用抗CD3抗体与7-AAD（7-氨基放线菌素D）进行染色。在CD3+7AAD-细胞群中设门后，利用计数微球对CFSE稀释细胞的数量进行计数。本实验以复孔开展，生物学重复次数n=4。