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. 2014 Apr 15;25(8):1216–1233. doi: 10.1091/mbc.E13-08-0471

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© 2014 Kasioulis et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 4: — Kdm3a encodes two protein isoforms. (A) Intron-exon diagram of Kdm3a. Short isoform i2 was cloned following ENSEMBL prediction (ID numbers between parentheses). First and unique Kdm3a-i2 exon is contained within intron 11. (B) Diagram illustrates the amino acid position of Kdm3a protein domains in full-length (FL) and isoform 2 (i2). Green line, i2-exclusive exon; amino acid sequence shown in bold. pAb indicates the relative position of amino terminal Kdm3a single polyclonal antibody used along this study. (C) qRT-PCR from testis and MEF RNA show a decrease of Kdm3a-FL and unaltered transcript levels of i2 in Kdm3a^GT/GT testis. A slight reduction of Kdm3a-i2 is observed in Kdm3a^GT/GT primary MEFs. (D) RPE1 cells transfected with GFP tag fused upstream of Kdm3a-i2. Monoclonal antibodies to mono- or dimethylated Lys-9 of histone H3 were used to determine the state of histone methylation (red) in transfected cells (green). Arrows point to transfected cells. (E) Quantitation of (D). n = number of cells counted for each condition. *p < 0.05, Fisher's exact test.