Fig. 1.
“Reverse northern” analysis of IEG induction by seizure. Duplicate Southern blots were probed with radiolabeled cDNA prepared from hippocampus of naive control (A) and MECS with cycloheximide-stimulated (B) rats. β-Tubulin and GAPD are constitutively expressed and provide a comparison for genes that are induced by MECS. Comparison of blots A andB demonstrate that MECS increases the intensity of hybridization to zif268, c-jun, and c-fos. C, Blots demonstrate a conventional Northern analysis of 2 μg Poly(A+)RNA prepared from hippocampus of naive control rats, rats treated with cycloheximide for 3 hr (3Hr CHX), or cycloheximide in combination with MECS (CHX/MECS) and probed with zif268, c-fos, andGAPD. The sensitivity of the reverse Northern is comparable to conventional Northern methods and permits analysis of multiple genes from small tissue samples.