1. Volume 12, Issue 6, Pages 1591-1598 (December 2003)
Histone Methyltransferases Direct Different Degrees of Methylation to Define Distinct Chromatin Domains 
Judd C. Rice, Scott D. Briggs, Beatrix Ueberheide, Cynthia M. Barber, Jeffrey Shabanowitz, Donald F. Hunt, Yoichi Shinkai, C.David Allis 
Molecular Cell 
Volume 12, Issue 6, Pages (December 2003)
DOI: /S (03)

2. Figure 1
Characterization of the H3 Lys9 Mono-, Di-, and Trimethyl-Specific Antibodies
(A) The first 30 amino acids of histone H3 are shown. Human H3 lysine residues known to be posttranslationally modified are depicted: red for methylation, green for acetylation, and yellow for both. Red lollipops indicate that H3 Lys9 can be mono-, di-, or trimethylated. The purple line from residues 5–16 denotes the branched synthetic peptide used for immunization. The shaded box reflects the epitope similarity between Lys9 and Lys27.
(B) Western analysis of the antibodies using unmodified recombinant H3 (rH3), chicken histones, and human HeLa and mouse fibroblast whole-cell lysates. Each antibody only detects modified H3 (arrow).
(C) Peptide competition, followed by Western analysis on HeLa whole-cell lysates, using the different Lys9 methyl-specific antibodies with synthetic peptides that were unmodified, mono-, di-, or trimethylated at Lys9 or Lys27. Only the appropriate corresponding peptide can eliminate the signal for each of the antibodies.
(D) Peptide competition, followed by Western analysis on HeLa whole-cell lysates, using the different Lys9 methyl-specific antibodies with methylated synthetic peptides for H3 Lys4, Lys36, Lys79, and acetylated Lys9. Only the mono-, di-, or trimethylated H3 Lys9 peptide (Lys9-methyl*) could absorb the signal of the mono-, di-, and trimethyl-specific antibodies, respectively.
Molecular Cell  , DOI: ( /S (03) )

3. Figure 2
Subnuclear Localizations of H3 Lys9 Mono-, Di-, and Trimethylation
(A) Immunofluorescence staining of mouse embryonic fibroblasts using the H3 Lys9 methyl-specific antibodies. Antibody staining is visualized as red, DNA staining with DAPI is visualized as green, and an overlap between the two is visualized as yellow when merged.
(B) Dual immunofluorescence staining was performed using the H3 Lys9 mono- (red) and dimethyl-specific (green) antibodies.
(C) Dual immunofluorescence staining with either the H3 Lys9 mono- or dimethyl-specific antibodies (red) and a phosphorylated RNA Pol II antibody (green).
Molecular Cell  , DOI: ( /S (03) )

4. Figure 3
G9a Directs H3 Lys9 Mono- and Dimethylation within Euchromatin
(A) Western analysis using the H3 Lys9 methyl-specific antibodies on lysates from wild-type mouse ES cells (WT), G9a heterozygous deletes (G9a+/−), G9a homozygous deletes (G9a−/−), and G9a−/− cells containing a wild-type G9a transgene (G9a−/− + G9a WT). Coomassie staining of core histones and blotting with a general H3 antibody was performed to confirm equal loading.
(B) Immunofluorescence staining in the G9a mouse ES cells using the H3 Lys9 methyl-specific antibodies.
Molecular Cell  , DOI: ( /S (03) )

5. Figure 4
Suv39h1 and Suv39h2 Direct H3 Lys9 Trimethylation at Pericentric Heterochromatin
(A) Western analysis using the H3 Lys9 methyl-specific antibodies on lysates from wild-type MEFs (WT), Suv39h1/Suv39h2 double null MEFs (DN), and DN cells infected with a FLAG-tagged wild-type retroviral construct of gfp, Suv39h1, or Suv39h2. Relative amounts of Suv39h1 and Suv39h2 protein in the infected cells were determined with a FLAG antibody. Blotting with an H3 Lys9-acetyl and Lys4-dimethyl antibody was used to assess global changes of these H3 modifications in the different cell types. Coomassie staining of core histones and blotting with a general H3 antibody was performed to confirm equal loading.
(B) Immunofluorescence staining in the WT, DN, and DN cells infected with Suv39h1 and Suv39h2 using the H3 Lys9 methyl-specific antibodies.
Molecular Cell  , DOI: ( /S (03) )