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25 A +1 -1787+19 hMLH1 B Acetylated H3 Enrichment SW480 RKO Position from +1 BBII ++-- SW480 BIBI ++-- RKO C hMLH1 GAPDH Ab D Methyl-H3-K4 enrichment Position.

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Presentation on theme: "25 A +1 -1787+19 hMLH1 B Acetylated H3 Enrichment SW480 RKO Position from +1 BBII ++-- SW480 BIBI ++-- RKO C hMLH1 GAPDH Ab D Methyl-H3-K4 enrichment Position."— Presentation transcript:

1 25 A hMLH1 B Acetylated H3 Enrichment SW480 RKO Position from +1 BBII ++-- SW480 BIBI ++-- RKO C hMLH1 GAPDH Ab D Methyl-H3-K4 enrichment Position from +1 SW480 RKO SW480 BBII ++-- RKO BIBI ++-- E GAPDH hMLH1 Ab Figure 1 F Methyl-H3-K9 enrichment Position from +1 SW480 RKO GAPDH hMLH1 Ab SW480 BBII ++-- BIBI ++-- G RKO

2 27 Figure 2 Methyl-H3-K4 enrichment BBBBIIII TSA Ab hMLH1 GAPDH C Position from +1 D -TSA +TSA Methyl-H3-K9 enrichment Position from +1 E F hMLH1 GAPDH BBBBIIII TSA Ab TSA +TSA Acetylated H3 enrichment A GAPDH hMLH1 BBBBIIII TSA Ab Position from +1 B -TSA +TSA

3 29 Figure 3 Position from Aza-dC +5-Aza-dC E Methyl-H3-K9 enrichment F hMLH1 GAPDH 5-Aza-dC Ab BBII BBII Aza-dC +5-Aza-dC Position from +1 C Methyl-H3-K4 enrichment D GAPDH 5-Aza-dC Ab BBII BBII hMLH1 B BBII GAPDH 5-Aza-dC Ab BBII Position from +1 A Acetylated H3 enrichment -5-Aza-dC +5-Aza-dC

4 31 Figure 4

5 33 +- RT GAPDH hMLH1 mock 5-Aza-dC mock 5-Aza-dC dH 2 O 12 h24 h48 h5 d B mock UMUUUUUUUUUUUMMMMMMMMMMM 5-Aza-dC mock 5-Aza-dC mock RKO SW480 PCR dH 2 O Bisulfite dH 2 O 12 h24 h48 h5 d A Figure 5

6 53 β-Actin H3.3-HA WT β-Actin H3.3-HA B A 18 bp linker 408 bp90 bp H3.33xHA Clone 10 Clone 16 WT DAPIAnti-HAMerge C D RKO 2 β-Actin H3.3-HA Figure 6

7 55 Figure 7 B RKO WT β-Actin H3.3-HA β-Actin H3.3-HA A RKO WT β-Actin H3.3-HA C β-Actin H 3.3-HA RKO WT SW480 WT SW480 clone 20 RKO Clones

8 56 Figure IgG RKO WT RKO Clones SW480 clone 20 SW480 WT RKO WT RKO Clones Figure 8. H3.3-HA can be immunoprecipitated with anti-HA. Whole cell (top panel) or nuclear (bottom panel) lysates were immunoprecipitated with an anti-HA antibody. Numbers across the top indicate clone number. Whole cell lysate from RKO clone 18 was incubated with normal rabbit IgG as a negative control.

9 58 Figure 9 B Position from +1 Methyl-H3-K4 Enrichment SW480 clone SW480 WT RKO clone RKO WT A H3.3 Enrichment Position from +1 SW480 clone SW480 WT RKO clone RKO WT


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