Materials and Reagents

1. Sterile Syringe Filters (Merck Millipore Corporation, Millex, catalog number: SLGV033RS )
   
2. Ice
   
3. Seeds of A. thaliana ecotype Columbia (Col-0)
   
4. Escherichia coli (BL21) (Thermo Fisher Scientific, Invitrogen^TM, catalog number: C6000-03 )
   
5. pGEX-4T-3 expression vector (GE Healthcare, Amersham, catalog number: 28-9545-52 )
   
6. Agrobacterium (GV3101)
   
7. LB medium (Caisson Laboratories, catalog number: LBP01-500 GM )
   
8. Murashige and Skoog (MS) media (Sigma-Aldrich, catalog number: M5524 )
   
9. Ampicillin (Beyotime, catalog number: ST007 )
   
10. Isopropyl-b-D-thiogalactopyranoside (IPTG) (Sigma-Aldrich, catalog number: 367-93-1 )
    
11. Sucrose (Sigma-Aldrich, catalog number: 57-50-1 )
    
12. Potassium hydroxide (KOH) (Sigma-Aldrich, catalog number: 1310-58-3 )
    
13. Bacto Agar (Sigma-Aldrich, catalog number: 9002-18-0 )
    
14. Ethanol
    
15. NP40 (Sigma-Aldrich, Abcam, catalog number: 9016-45-9 )
    
16. Sodium chloride (NaCl) (Sigma-Aldrich, catalog number: 31434 )
    
17. Potassium chloride (KCl) (Sigma-Aldrich, catalog number: 7447-40-7 )
    
18. Sodium phosphate dibasic (Na₂HPO₄) (Sigma-Aldrich, catalog number: 7558-79-4 )
    
19. Potassium phosphate monobasic (KH₂PO₄) (Sigma-Aldrich, catalog number: 7778-77-0 )
    
20. Hydrochloric acid (HCl)
    
21. Ethylenediaminetetraacetic acid (EDTA) (Sigma-Aldrich, catalog number: 60-00-4 )
    
22. Glycerol (Sigma-Aldrich, catalog number: 56-81-5 )
    
23. L-Glutathione reduced (Sigma-Aldrich, Abcam, catalog number: 70-18-8 )
    
24. Ultrapure water
    
25. Liquid N₂
    
26. GST^.Bind^TM Resin (Merck Millipore Corporation， Novagen, catalog number: 70541 )
    
27. Protease inhibitors (Roche Diagnostics, catalog number: 11873580001 )
    
28. Glycine (Sigma-Aldrich, catalog number: 15527 )
    
29. Tris base (Sigma-Aldrich, catalog number: 77-86-1 )
    
30. GFP-Trap (ChromoTek, GFP-Trap^®_M)
    
31. HDAC activity colorimetric assay kit (BioVision, catalog number: K331-100 )
    
32. HeLa nuclear extracts (Biovision, catalog number: 1641-1 )
    
33. Trichostatin A (Sigma-Aldrich, catalog number: 58880-19-6 )
    
34. Bradford Protein Assay Kit (Beyotime, catalog number: P0006 )
    
35. Bovine Serum Albumin (BSA) (Sigma-Aldrich, catalog number: A7906 )
    
36. Phenylmethanesulfonyl fluoride (PMSF) (Sigma-Aldrich, catalog number: P7626 )
    
37. Boc-Lys(Ac)-pNA
    
38. Phosphate buffered saline (PBS) (see Recipes)
    
39. GST Elution buffer (see Recipes)
    
40. Plant protein extraction buffer (see Recipes)
    
41. GFP Wash buffer (see Recipes)
    
42. GFP Elution buffer (see Recipes)
    
43. HDAC Assay Buffer (Biovision) (see Recipes)
    
44. HDAC substrate (colorimetric substrate) (see Recipes)
    

Equipment

1. Plant growth chamber (Panasonic Healthcare Corporation, model: MLR-352 )
   
2. Sterile fume hood (Alibab, Airtech, model: VS-1300L )
   
3. Autoclave (HIRAYAMA, model: HVE-50 )
   
4. Centrifuge (Eppendorf AG, model: 5418R and 5810R )
   
5. Shaker (Eppendorf AG, model: New Brunswick^TM Innova^® 40 )
   
6. Sonicator (SONICS & MATERIALS, model: VCX130 )
   
7. Spectrophotometer (Molecular Devices Spectra Max)
   

Procedure