Additional file 3: of Human central nervous system astrocytes support survival and activation of B cells: implications for MS pathogenesis

Figure S2. Effects of astrocytes cytokine neutralization on B cell survival and activation. B cells from HC were either cultured alone, or with stimulated astrocyte conditioned-medium (ACM), or with ACM pre-treated with neutralizing antibodies to IL-6 (a, b; anti-IL6: aIL-6), IL-15 (c, d; anti-IL-15: aIL-15) or BAFF (e, f; anti-BAFF: aBAFF); or pre-treated with corresponding isotype control antibodies. After 2 days of culture B cell viability was assessed using ANNEXIN V and 7AAD staining, and CD86 expression was measured by flow cytometry (representative experiment). (TIFF 4226 kb

Additional file 2: of Human central nervous system astrocytes support survival and activation of B cells: implications for MS pathogenesis

Figure S1. Confirming activation of human astrocytes. Astrocytes were cultured for 24 h and were either left unstimulated or were stimulated with IFNγ (10 ng/ml) and IL-1β (10 ng/ml). After 24 h, the astrocytes were washed thoroughly and fresh medium was added. After an additional 24 h in culture, at which time cultures were imaged and supernatants were collected for subsequent measurement of astrocyte-secreted IL-6 by ELISA. Compared to unstimulated astrocytes (a), stimulated astrocytes exhibited activated morphology (b) and significantly-enhanced production of IL-6 (c; p = 0.0016; paired t-test). (TIFF 3951 kb