Figure 2.

RT-PCR analysis of TNF-α gene expression in rat neonatal microglia after in vitro treatment with LPS or DOM. Neonatal rat brain microglia (2.8–5.0 x 10⁶ cells/culture dish) were treated with (A) LPS [3 ng/mL] or (B) DOM [1mM] for 4 to 24 hours as described in Experimental. Amplification of TNF-α and S12 mRNAs shows the predicted cDNA size after separation on a 1.5 % agarose gel and visualization by SYBR^R Gold nucleic acid staining. The S12 ribosomal RNA gene product was amplified for 25 and 30 cycles and demonstrated equal loading of the gels. Quantification of the TNF-α product was done after 25 cycles, where mRNA expression was observed to be in the linear range. The figure depicts one of two similar experiments.