JCS019943 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 -

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• Supplemental Table S2 -

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• Supplemental Figure S1 -

  Fig. S1. Mutant xpo1-1 inhibits nuclear protein export of a control protein. BMA38a (WT) and MPS15 (xpo1-1 LSM7-13myc) with pKW430 (NLS-NES-GFP) were grown in SD-Ura-Met at 23°C and shifted to 37°C for 15 minutes, before fixing the cells. NLS-NES-GFP is shown in green, DAPI in red.

• Supplemental Figure S2 -

  Fig. S2. Lsm7p and Lsm8p localization is not affected by depletion of SmD1p. Cultures of MPS7, MRY76 and MRY77 were grown at 30°C in YPGalA to log phase and then for a further 12 hours in YPDA, while keeping the cells in log phase. (A) Localization of 13-myc-tagged Lsm7p or Lsm8p (shown in red) before (Gal) and after (Glu) growth in YPDA. (B) Total RNA isolated from the same cultures before and after growth in glucose was analyzed by northern blotting, probing for U3A, U1, U6 and scR1 RNAs.

• Supplemental Figure S3 -

  Fig. S3. Lsm8-GFP is de-localized from the nucleus after hyperosmotic shock, but recovers after longer periods of time. MPS11 (P_GAL-LSM8) with pMR83 (Lsm8-GFP) was grown in SD-Ura-Met and shifted to SD-Ura-Met with 0.6 M or 1 M NaCl, or 1 M KCl or 1 M sorbitol only. Localization of Lsm8-GFP was examined in live cells after 5, 15, 30 and 120 minutes of incubation.

• Supplemental Figure S4 -

  Fig. S4. Recovery after hyperosmotic shock is independent of Hog1p protein kinase. MRY83 (hog1Δ LSM8-13myc) cells were grown in YPDA and shifted to YPDA with 0.6 M NaCl. Cells were fixed after 5, 15, 30, 60 and 120 minutes of incubation. Some cells were shifted back to YPDA (and incubated for 60 or 120 minutes) after incubation in YPDA with 0.6 M NaCl for 15 minutes. Lsm8-13myc is shown in red, DAPI in green.