Fig. 2

IFNγ-regulated expression and function of PD-L1 in astrocytes. A Primary human spinal cord astrocytes and B primary murine spinal cord astrocytes were stimulated with and without 10 ng/ml IFNγ for 24 h and RNA was collected and analyzed for transcript levels of CD274 and PDCD1 by qRT PCR. C Primary human spinal cord and D primary murine spinal cord astrocytes were stimulated with or without 10 ng/ml IFNγ for 24 h and protein lysates were assessed for expression of PD-L1 via Western blot. E, F PD-L1 levels were quantified and normalized to β-actin expression. G Schematic representation of the experimental design for data presented in H. H Caspase 3/7 activity was quantified and normalized to lymph node cell number following coculture with astrocytes treated with 10 ng/ml IFNγ. Data are representative of 2 independent experiments with 3–4 technical replicates each. All data represent the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001 compared to media-treated samples and ^††P < 0.01, ^†††P < 0.001 between treatments by two-way ANOVA