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. 2012 Jan 31;22(6):1058–1077. doi: 10.1038/cr.2012.20

Figure 4.

Figure 4

Requirement for Rts1, Rrd1 and Rrd2 in glucose-induced activation of PP1 and interaction of Rts1 with PP2A and PP1. At time 0, 20 mM glucose was added to glucose-deprived (glycerol-grown) cells of BY-strains with different deletions and cell extracts were used to measure specific PP1 activity. (A) WT (•) and pph21Δ pph22Δ (○). (B) WT strain (•), cdc55Δ (○) and rts1Δ (▴). (C) pph21Δ pph22Δ (•) and rts1Δ pph21Δ pph22Δ (○). (D) WT (•), rrd1Δ (○), rrd2Δ (▴) and rrd1Δ rrd2Δ (△). (E) WT cells expressing HA-tagged Pph21 or Glc7 were grown to exponential phase in either glucose (YPD) or glycerol (YPGly). A third sample was taken 2 min after addition of 20 mM glucose to the glycerol-grown cells (YPGly + Glu). Antibodies used for the immunopurification (IP) and immunodetection (western blot): HA: anti-HA; Rts1: anti-Rts1; -: no antibody. (F) Detection of Glc7 and Pph21 in total extract and in extract IP using the indicated antibodies by means of western blot with the antibodies indicated as follows: HA: anti-HA; Rts1: anti-Rts1; Pph21: anti-Pph21; -: no antibody. Strains used are: (1) WT + pHA-Glc7, (2) WT + pHA-Pph21, (3) rts1Δ + pHA-Pph21 and (4) pph21Δ pph22Δ + pHA-Glc7.