Skip to main content
. 2017 Mar 1;8(16):26802–26818. doi: 10.18632/oncotarget.15818

Figure 7. SET may modulate gene expression through hypoacetylation of histones and increase of HDAC1 levels.

Figure 7

(A) Western blot assay was performed by adding antibodies against acetyl-H2B, acetyl-H3, SET, and β-actin to total proteins extracted from HEK293, HEK293/SET, HN12, HN12siSET, HN13, and HN13siSET cells. (B) HDAC1 levels were assessed using immunofluorescence assay. Cells from five fields were measured using ImageJ software. Graphics represent the percentage of TET1-positive cells. Red arrows indicate SET accumulation; green arrows indicate SET downregulation. (C) To assess whether SET directly associates with sites of hypoacetylated gene promoter, HEK293 and HEK293/SET cells were treated with the histone deacetylase inhibitor Trichostatin A (TSA; 100 ng/mL) for 24 h. After cell lysis, chromatin was immunoprecipitated with anti-SET antibody and PCR was performed using primers that recognize the promoter of HIF-1A and NFATC3. INPUT samples consist of total DNA; SET lanes refer to DNA immunoprecipitated with anti-SET antibody. (D) Western blot assay using antibodies against acetyl-H2B, acetyl-H3, SET, and β-actin was performed with total proteins from the respective cells treated with TSA (100 ng/mL) for 24 h.