Figure 4.
Irc21 Inhibits DDR through PP2A Activation
(A) Cells were treated with 200 ng/mL rapamycin. Left: band-shift assays following phosphorylation of the PP2A branch proteins Gln3, Nnk1, Npr1, and Rtg3 after 30 min of rapamycin treatment. A horizontal line has been overlaid to assist in determining mobility shifts. Right: band-shift assays following the phosphorylation of Sch9 after 2 hr of rapamycin treatment.
(B) WT and irc21Δ cells were grown on YPD plates with or without rapamycin, metformin, caffeine, and wortmannin (left). All drugs are inhibitors of the Torc1-Tap42 pathway, represented at the right.
(C, E, and F) Cells were grown on YPD plates with or without 3 mM HU.
(D) Cells were arrested with α-factor and released in YPD with or without 0.2 M HU. Cells were treated for 3 hr and harvested to detect Rad53.
(G) Cells were arrested in G1 with α-factor and released in YPD with 0.2 M HU. After 3 hr, cells were released into YPD. Samples were collected at the indicated times to detect Rad53.
(H) Cells were grown on YPD + HU plates with or without okadaic acid (OA).
See also Figure S4.