Figure 2. Stm1 forms vacant 80S ribosomes upon TORC1 inhibition.
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A–DPolysome profiles of wild‐type (wt) and stm1Δ cells with (A–C) or without (D) CHX treatment under nutrient‐rich conditions (A, D), with rapamycin treatment for 1 h (B), or under nitrogen starvation for 1 h (C) using 5–50% sucrose density gradients containing 150 mM KCl. At least three biological replicates are analyzed for each condition.
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E, FPolysome profiles of formaldehyde cross‐linked wild‐type (wt) and stm1Δ cells using 5–50% sucrose density gradients containing 150 mM KCl (E) or 300 mM KCl (F).
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GPolysome profiles of stm1Δ cells containing either full‐length or N‐terminally deleted or C‐terminally deleted Stm1 mutants upon nitrogen starvation for 1 h.
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HBinding of full‐length or deletion mutants of Stm1 across the polysome profile fractions shown in Fig 2E. Nine equal‐volume fractions of each polysome profile were precipitated and analyzed by immunoblot for Stm1 and Rpl3. At least two biological replicates are used for each mutant.