Abstract

<p>A) Top panel shows the structure of WHSC1 gene. The position of the different translocation events in MM is shown by slashed lines. Grey and black ovals indicate non-coding and coding exons respectively, dotted straight line between exons represent the introns. Splicing event for the canonical longest MMSET isoform is shown by joining lines between exons. Exons 4a and 11, shown by white triangles, are used only for alternative splicing and are not present in MMSET II and REIIBP isoforms. Transcription start sites and first translated codons (ATG) for MMSET II and REIIBP are indicated respectively by bent black arrows and open arrows. Genomic position for ACA11 is shown by a white arrow. Bottom panel shows the protein structure of the two major isoforms of the whcs1 gene. The major domains of the proteins are, PWWP (Pro-Trp-Trp-Pro motif) a chromatin associated domains, HMG high mobility group, PHD plant homeo domain, SET set domain with pre and post motifs. The position of the nucleolus exclusion signal and nucleus localization signal sequences are based on published observations <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099493#pone.0099493-Keats2" target="_blank">[15]</a> and prediction analysis software <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099493#pone.0099493-Brameier1" target="_blank">[58]</a>. B) IF and confocal analysis on HeLa::REIIBP and HeLA::ΔSET. REIIBP-Flag (green) localizes in the cytoplasm and nucleolus in HeLa cells overexpressing REIIBP-Flag. Single nucleolus zoomed in as inset in each frame. C) Western blot analysis on nuclear extracts. The antibody used is indicated on the left. D) Percentage of Apoptotic cells in the shown line. Error bars come from the SEM (standard error of the mean) of 3 different analysis. E) Percentage of cells at different stages of the cell cycle. Each data is the average of three analysis. * p value <0.05. F) WST-1 proliferation assay for the indicated cells. Error bars are the SEM of 5 different analysis. C–F) The control is the empty lentivector transduced into HeLa. See also <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0099493#pone.0099493.s001" target="_blank">Figure S1</a>.</p

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