Fig. 6. Rgf3 is independent of the SIN. (A) The rgf1::ura4⁺ strain (KGY 5385) was grown at 32°C in YE, fixed with ethanol, and stained with DAPI. (B) pARTGFP-Rho1 was transformed into lad1-1 cells. Transformants were grown at 25°C, filtered and resuspended in YE medium and shifted to 36°C for 4 hours. GFP-Rho1p localization was visualized in live cells. Arrowheads indicate medially localized GFP-Rho1p. (C) rgf3-GFP sid2-250 cells (KGY5326) were synchronized in G2 phase by lactose gradient centrifugation, filtered, resuspended in fresh medium at 36°C and followed throughout a cell cycle. The percentage of binucleate (anaphase) cells, septated cells and cells with Rgf3p-GFP rings were determined every 20 minutes.