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Figure S1. Mixed neuron-glia cultures contain a small proportion of endothelial cells. Representative images of mixed neuron-glia cultures stained for microglia (CD45+Iba1+), neurons (MAP2+NF200+), astrocytes (GFAP+), OPCs (Olig2+ABA5+PDGFRα+Nkx2.2+), oligodendrocytes (Olig2+O4+APC+PLP+MBP+, endothelial cell (CD31+), pericytes (PDGFRβ+) and NPCs (Nestin+Sox1+). Figure S2. Single exogenous Treg-polarizing factors do not affect OPC differentiation in mixed neuron-glia cultures. Mixed neuron-glia cultures were treated with (A) IL-2 (500 pg/ml) or (B) TGF-β (300 pg/ml) or anti-IFN-γ (500 ng/ml) compared to Treg-conditioned media and matched medium control (BSM) for 5 days. Immunofluorescence analysis of MBP+ area, n = 6 wells, mean +/− SEM, unpaired t-test, representative of two independent experiments. FOV = field of view = 1 mm2. Figure S3. NeuroBrew-21 negatively affects OPC differentiation and cell survival. Mixed neuron-glia cultures were treated with normal differentiation medium (= + NeuroBrew-21) or with differentiation medium without NeuroBrew-21 (= -NeuroBrew-21) for 5 days. Immunofluorescence analysis of EthD-1+ cell counts and MBP+ area, n = 6 wells, mean +/− SEM, unpaired t-test, representative of two independent experiments. FOV = field of view = 1 mm2. (PPTX 5757 kb) |
