Journal: Nucleic Acids Research

Article Title: H2A.Z.2.2 is an alternatively spliced histone H2A.Z variant that causes severe nucleosome destabilization

doi: 10.1093/nar/gks267

Figure Lengend Snippet: Identification of Z.2.2. ( A ) qPCR with cDNA from different human cell lines and tissues using primers specific for Z.2.1 and Z.2.2. Data were normalized to HPRT1 and HMBS expression levels. Controls generated without reverse transcriptase (no RT) were used to assess amplification threshold. Shown are the levels of Z.2.2 mRNA as percentages of total Z.2 transcripts (Z.2.1 + Z.2.2). For an evaluation of absolute expression levels see Supplementary Figure S1C and D . ( B ) Amino acid alignment of human Z.1, Z.2.1 and Z.2.2 proteins using ClustalW Alignment (MacVector 10.0.2). Identical amino acids are highlighted in dark gray, similar amino acids in light gray and changes are set apart on white background. Known acetylation sites are depicted with stars and ubiquitination sites with circles. A schematic representation of the secondary structure of Z.1 and Z.2.1 is shown below the alignment, including depiction of the H3/H4 docking domain . M6 and M7 boxes indicate regions important for H2A.Z-specific biological functions in D. melanogaster . ( C ) IB analyses of RP–HPLC purified fractions from different human (HEK293, HeLa, HeLa Kyoto and U2OS) and mouse (NIH3T3) cell lines using a polyclonal rabbit αZ.2.2 and αH2A.Z (αZ, C-terminal) antibodies. Recombinant Z.2.2 protein (rZ.2.2) was loaded in the first lane as positive control for αZ.2.2 antibody. Similar results were obtained when using a monoclonal rat αZ.2.2 antibody (data not shown).

Article Snippet: For an evaluation of absolute expression levels see Supplementary Figure S1C and D . ( B ) Amino acid alignment of human Z.1, Z.2.1 and Z.2.2 proteins using ClustalW Alignment (MacVector 10.0.2).

Techniques: Expressing, Generated, Reverse Transcription, Amplification, Ubiquitin Proteomics, Purification, Recombinant, Positive Control