Figure 4

RT-PCR transcript analysis of TNF-α gene expression. Neonatal rat brain microglia (2.8–5 × 10⁶ cells/culture dish) were treated with (A) LPS [10 ng/mL] or (B) Domoic acid [1 mM] for 1 to 6 hours. (See Materials and Methods). Amplification of TNF-α and S12 genes shows the predicted fragment size after separation on a 1.5 % agarose gel and visualization by SYBR^R Gold nucleic acid staining. The S12 rRNA gene product was amplified for 25 cycles and demonstrates equal loading of the gels. Quantification of the TNF-α product was done after 20 cycles (LPS) or 25 cycles (Domoic acid). The cycle numbers used for quantification were shown to be in the linear range.