A) FLAG tagged dRif1 is expressed in S.cerevisiae. Total cell extracts of yeast transformed with empty vector (lane 1) or 3XFLAG tagged dRif1(lane 2) were probed with antibodies to FLAG and tubulin. Lanes 2 shows 3XFLAG dRif1 expression. B) dRif1 introduced in yeast localizes to the nucleus but not to yeast telomeres. Yeast cells transformed with empty vector or 3X FLAG dRif1 were stained for 3xFLAGdRif1 (green) and 13xmycSir4 (red). Nuclei were stained with DAPI (blue; Panel B, row 1&2). DNA damage induction with 0.05%MMS for 90 minutes treatment did not alter the localization of dRif1 in yeast (Panel B, rows 3,4). (Scale bar, 5 μm).