Fig. 5. Effect of gamone 2 on the promotion of gamone 1 transcription. (A) We examined the transcriptional levels of gamone 1 in type I cells in the early stationary phase after the addition of gamone 2 (G2). Total RNA was prepared and separated (6 µg/lane) from cells that had been incubated with gamone 2 for 0, 2, 4, 6, 8 or 20 hours, and gamone 1 mRNA was detected by northern hybridization using probe 2. A peak of transcriptional activity was reached in cells stimulated for 4-6 hours with gamone 2. The graph indicates the relative amounts of gamone 1 mRNA. (B) SDS-PAGE analysis and (C) biological activity tests of CFFs collected from individual cell suspensions.