Fig. 2

Embryonic stem cell-derived astrocytes undergo activation in response to a neuroinflammatory stimulus. We used the c-Jun N terminal kinase (JNK) pathway, a three-tiered mitogen-activated protein kinase pathway (MAPK) [95], to evaluate neuroinflammatory stimulation of ES-derived astrocyte-like cells by assessing the phosphorylation of c-Jun at either the Serine site 63 or 73. We exposed our cells to cytokines and then evaluated the JNK pathway (A) after both 2 h following cytokine stimulation (B, C), and 24 h after cytokine stimulation (D, E). At 2 h, P–c-Jun was increased in treated groups (F), while GLT-1 expression was similar across treatment groups (G), indicative that the maturity state of the cells was not altered, and that the astrocyte-like cells were afflicted by this stimulus (H). Congruent findings were seen after 24 h of cytokine stimulation (I-K). Significance level: NS, non-significant; *, p ≤ 0.05; **, p ≤ 0.01, ***, p < 0.001. Abbreviations: ES, embryonic stem cells; GLT-1, glutamate transporter 1; IL-1α, interleukin 1α; JNK, c-Jun N-terminal kinase; MAPK, mitogen activated protein kinase; P–c-Jun, phosphorylated c-Jun; Ser, serine; TNF-α, tumor necrosis factor α. Scale bars: B, D: 100 μm; C, E: 25 μm