Figure 8.

Leukemia inhibitory factor (LIF) secretion in primary astrocytes is constitutive and independent of NECA stimulation. (A) illustrates LIF immunocytochemistry in cultured cortical astrocytes, where LIF is found in vesicle-like structures throughout the cytoplasm. Scale bar corresponds to 10 μm. (B) shows complete co-localization of LIF with Giantin, a marker of Golgi apparatus, when cultured astrocytes were treated with Brefeldin A (BFA, 5 μg/mL for 1 hour). Scale bar corresponds to 10 μm. (C) shows a representation of sandwich ELISA experiment performed to detect LIF content in supernatants of cultured astrocytes that were treated without or with NECA (1 μM) for 4 hours. Where indicated, cells were pre-treated with BFA (5 μg/mL) for 1 hour prior to NECA stimulation. Each bar corresponds to the mean concentration of LIF in triplicate samples; error bars indicate SEM. Observations were confirmed by repeating the experiments two additional times. P < 0.05. NECA, 5′-N-ethylcarboxamide.