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. 2016 Jun 3;291(31):16231–16239. doi: 10.1074/jbc.M115.713206

FIGURE 1.

FIGURE 1.

Nitrogen starvation elevates Ras1 phosphorylation. A, whole cell extracts from wild-type cells and ras1Δ cells were resolved by 8% SDS-PAGE and probed with anti-Ras antibodies (ab79973, Abcam). Equal loading of each lane was confirmed via immunoblotting (IB) with anti-Pgk1. p-Ras1, phosphorylated Ras1; Ras1, unphosphorylated Ras1. B, wild-type cells (BY4741) were grown to mid-log phase in yeast extract peptone dextrose (YPD) medium (time 0) and continued to grow for the indicated times. Whole cell extracts were resolved by 8% SDS-PAGE and probed with anti-Ras antibodies. Equal loading of each lane was confirmed via immunoblotting with anti-Pgk1. Quantification of immunoblots by densitometry from three independent experiments is shown in the right panel. The difference between each time point and time 0 was statistically analyzed (*, p < 0.050). C and D, wild-type cells (BY4741) were grown in YPD medium to mid-log phase and then switched to medium without glucose (w/o glucose, C) or without nitrogen (w/o nitrogen, D) for the indicated times. Whole cell extracts were resolved by 8% SDS-PAGE and probed with anti-Ras antibodies. The data shown are representative of three independent experiments. Quantification of immunoblots by densitometry from three independent experiments is shown in the right panels. The difference between each time point and time 0 was statistically analyzed (**, p < 0.010).