1. 《进展》稿件标题：环境变化和时序衰老过程中酵母蛋白激酶 Sch9 磷酸化的调控 稿件编号： 20130025 刘军 闾磊 郄蓓蓓 刘科 （四川大学生命科学学院生物资源与生态环境教育部重点实验室 成都 610064 ） Western blot 全胶图 - 补充材料

2. Band of T570 site phosphorylated Sch9 sch9Δ-pRS416-SCH9-3HA 2% glucose 0.5% gluocse sch9Δ-pRS416-SCH9-3HA 0.5% glucose sch9Δ-Vector sch9Δ-pRS416-SCH9-3HA 2% glucose sch9Δ-pRS416-SCH9-3HA 0.5% glucose sch9Δ-Vector

3. Fig1 (a) Specificity of anti-T737-P antibody Sch9- 3HA Anti-HA Anti-T737-P T737-P Rapamycin: - + Sch9 T737 residue phosphorylation Non-specific band

4. Fig1 (b) Sch9-3HA (Stationary phase) Sch9- 3HA Time (min) 0 30 60 30 60 30 60 30 60 30 60 30 60 30 60 Non-specific band - C 2%Glc 2%Gal 2%Suc2%Mal2%Gal2%EtOH Anti-HA

5. Fig1 (b) T570-P (Stationary phase)

6. Fig1 (b) T737-P (Stationary phase) Time (min) 0 30 60 30 60 30 60 30 60 30 60 30 60 30 60 - C 2%Glc 2%Gal 2%Suc2%Mal2%Gal2%EtOH T737-P Non-specific band Anti-T737-P

7. Fig1 (c) Sch9-3HA (Exponential phase) Time (min) 0 10 30 10 30 10 30 10 30 10 30 ＋ Glc ＋ Gln － Glc ＋ Gln re-add Glc ＋ Glc － Gln re-add Gln Sch9-3HA Exponential phase(15h) T570A-Sch9- 5HA 24H Non-specific band Anti-HA

8. Fig1 (c) T570-P (Exponential phase) T570-P Anti-T570-P

9. Fig1 (c) T737-P (Exponential phase) This is a control that Sch9 T737 residue lose phosphorylation completely after glucose depletion Anti-T737-P

10. Fig1 (d) Sch9-3HA (Stationary phase) Time (min) 0 10 30 10 30 10 30 10 30 10 30 ＋ Glc ＋ Gln － Glc ＋ Gln re-add Glc ＋ Glc － Gln re-add Gln Sch9-3HA Early stationary phase(72h) T570A-Sch9- 5HA 72H Anti-HA

11. Fig1 (d) T570-P (Stationary phase) T570-P Anti-T570-P

12. Fig1 (d) T737-P (Stationary phase) This is a control that Sch9 T737 residue lose phosphorylation completely after glucose depletion Anti-T737-P

13. Fig2 (a) (Exponential phase) Long exposure time Short exposure time

14. Fig2 (b) Sch9-3HA (Exponential phase) Time (min) 0 10 30 60 10 30 60 10 30 60 10 30 60 Mock0.5 M NaCl0.3 M H 2 O 2 38 ℃ Sch9- 3HA Non-specific band Anti-HA

15. Fig2 (b) T570-P (Exponential phase) Time (min) 0 10 30 60 10 30 60 10 30 60 10 30 60 Mock0.5 M NaCl0.3 M H 2 O 2 38 ℃ T570-P Non-specific band Anti-T570-P

16. Fig2 (b) T737-P (Exponential phase) Time (min) 0 10 30 60 10 30 60 10 30 60 10 30 60 Mock0.5 M NaCl0.3 M H 2 O 2 38 ℃ T737-P Non-specific band Anti-T737-P

17. Fig2 (b) Actin Loading control (Exponential phase) Time (min) 0 10 30 60 10 30 60 10 30 60 10 30 60 Mock0.5 M NaCl0.3 M H 2 O 2 38 ℃ Anti- Actin Actin Non-specific band

18. Fig2 (c) Sch9-3HA (Exponential phase) Time (min) 0 10 30 10 30 10 30 10 30 10 30 10 30 MockpH2.5pH6.0 pH2.7 100 mM HAc pH4.0 100 mM HAc pH6.0 100 mM HAc Sch9- 3HA Non-specific band Anti-HA

19. Fig2 (c) T570-P (Exponential phase) Non-specific band

20. Fig2 (c) T737-P (Exponential phase) Time (min) 0 10 30 10 30 10 30 10 30 10 30 10 30 MockpH2.5pH6.0 pH2.7 100 mM HAc pH4.0 100 mM HAc pH6.0 100 mM HAc T737-P Non-specific band

21. Fig2 (d) Sch9-3HA (Exponential phase) Time (min) 0 10 30 60 10 30 60 10 30 60 10 30 60 Mock1 M Glc1 M Gly1 M Sorbitol Sch9- 3HA Non-specific band

22. Fig2 (d) T570-P (Exponential phase)

23. Fig2 (d) T737-P (Exponential phase) Time (min) 0 10 30 60 10 30 60 10 30 60 10 30 60 Mock1 M Glc1 M Gly1 M Sorbitol T737-P Non-specific band

24. Fig4 (a) Sch9-6HIS Time-course experiment Sch9-6HIS Over-expression T570-P Non-specific band Anti-HIS Anit-T570-P Time of Induction (h) 35.5 41 46 60 66 70 84 94 116

25. Fig4 (e) Sch9-3HA Time-course experiment

26. Fig4 (e) T570-P Time-course experiment Time (h) 15 18 22 28 32 36 48 72 96 120 144 168 216 Non-specific band T570-P 2%Glucose T570-P 0.5%Glucose Non-specific band 10 uM PHS T570-P

27. Fig4 (e) T737-P Time-course experiment Time (h) 15 18 22 28 32 36 48 72 96 120 144 168 216 T737-P 2%Glucose 0.5%Glucose Non-specific band 10 uM PHS Anti-T737-P

28. Fig4 (e) Actin Loading control 2%Glucose 0.5%Glucose Time (h) 15 18 22 28 32 36 48 72 96 120 144 168 216 Actin 10 uM PHS Actin Anti-Actin

29. Fig4 (f) T570-P (TB50 wildtype strain) Anti-T570-P

30. Fig4 (f) T737-P (TB50 wildtype strain) Time (h) 15 18 22 28 32 36 48 72 96 120 144 168 216 15 T737-P 2%Glucose 0.5%Glucose 10 uM PHS Anti-T737-P

31. Fig4 (f) Actin Loading control (TB50 wildtype strain) Time (h) 15 15 18 22 28 32 36 48 72 96 120 144 168 216 15 2%Glucose 0.5%Glucose actin Anti-Actin