Fig 6. The double knockout cells of lam2, gtr2 and npr2 lack synthetic effects in growth defect, canavanine resistance, or nuclear localization of Gaf1 induced by nitrogen depletion.

(A, B) Δgtr2Δgtr1 cells, Δgtr2Δlam2 cells, Δgtr2Δnpr2 cells, Δnpr2Δnpr3 cells, Δnpr2Δlam2 cells lacked synthetic effects in growth defect. The indicated prototrophic wild-type cells (wt, KP5080), Δgtr2 cells (KP6571), Δgtr1 cells (KP6573), Δgtr2Δgtr1 cells (KP6684), Δlam2 cells (KP6578), Δgtr2Δlam2 cells (KP6590), Δnpr2 cells (KP6506), Δgtr2Δnpr2 cells (KP6588), Δnpr3 cells (KP6552), Δnpr2Δnpr3 cells (KP6683) and Δnpr2Δlam2 cells (KP6682) were spotted onto the indicated plates, and then incubated at 27°C on EMM plates for 4 days, on EMM plates containing canavanine for 5 days, and on EMM plates containg rapamycin or Torin for 4 days. (C) Δgtr2Δgtr1 cells, Δgtr2Δlam2 cells and Δgtr2Δnpr2 cells lacked synthetic effects in nuclear localization of Gaf1 induced by nitrogen depletion. Wild-type cells (wt, KP6488), Δgtr2Δgtr1 cells (KP6687), Δgtr2Δlam2 cells (KP6685) and Δgtr2Δnpr2 cells (KP6686) expressing Gaf1-YFP under the native Gaf1 promoter were grown to mid-log phase in EMM medium. Fluorescent images of Gaf1-YFP were then acquired before (0 min) or after nitrogen depletion (-NH₄Cl) for the indicated time. Scale bar, 10 μm.