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      Correction: E2F1 and TFDP1 Regulate PITX1 Expression in Normal and Osteoarthritic Articular Chondrocytes

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      The PLOS ONE Staff
      PLoS ONE
      Public Library of Science

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          Abstract

          The caption for Fig 1 is incorrectly displayed as the fourth paragraph of the Results section. The publisher apologizes for the error. Please see the correct Fig 1 caption here. 10.1371/journal.pone.0167530.g001 Fig 1 Critical regulatory regions in the PITX1 promoter govern its expression in chondrocytes. Human C28/I2 chondrocyte cells were cotransfected with either the empty pGL4 plasmid (luciferase reporter plasmid) or the pGL4 plasmids containing different regions of the PITX1 promoter combined with either the empty pBabe plasmid or pBabe plasmids expressing ER (estrogen receptor) fused to E2F1, E2F2, or E2F3 and induced with 4OH-tamoxifen (OHT) for 24 h. (A) The -3895/+61 PITX1 gene region contains regulatory elements capable of producing a luciferase signal that is not affected by OHT treatment. (B) Overexpression of E2F1, E2F2, and E2F3 produces a significant increase in the luciferase activity under the control of the -3895/+61 PITX1 gene region. (C) The proximal sequence of the PITX1 promoter contains several E2F1 binding sites, as predicted by MatInspector 8.0 software (Genomatix Software Suite). (D) Overexpression of E2F1 has variable effects on luciferase activity depending on the length of the transfected promoter region (-3895/+61; -374/+61; -279/+61; -214/+61; -169/+61; -84/+61). Except for the -84/+61 PITX1 gene region, all the other constructs are significantly activated by E2F1. (Fig 1A, 1B and 1D) Data represents mean and standard deviation of 3 independent experiments. Asterisks represent a significant increase in luciferase activity (Two-way ANOVA; Bonferroni post hoc: *p < 0.0001) compared with control cells.

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          E2F1 and TFDP1 Regulate PITX1 Expression in Normal and Osteoarthritic Articular Chondrocytes

          We previously reported a loss-of-PITX1 expression in patients suffering of knee/hip osteoarthritis (OA). Search for the mechanism underlying this event led us to discover that PITX1 repression was triggered by the aberrant nuclear accumulation of Prohibitin (PHB1), an E2F1 co-repressor, in OA articular chondrocytes. In the current study, we assessed in details the involvement of E2F transcription factors in regulating PITX1 expression. We also analyzed other genes that are similarly regulated by E2F in regard to osteoarthritis. The transcriptional regulation of the PITX1 promoter by E2F1 was analyzed with the luciferase reporter assay, and chromatin immunoprecipitation assays, which confirmed direct E2F1-PITX1 interactions. The probable binding sites for E2F1 in the PITX1 promoter were identified by DNA pulldown experiments. In silico and in vitro analyses show that the PITX1 proximal promoter region contains 2 specific sequences that are bound by E2F1. Overexpression of E2F1 enhances PITX1 promoter activity and mRNA transcription. In primary control and osteoarthritis chondrocytes, real time RT-PCR was used to measure the mRNA expression levels of candidate genes under E2F1 transcriptional control. Transcription Factor Dp-1 (TFDP1) knockdown experiments confirmed that the E2F1-TFDP1 complex regulates PITX1. Knockdown of TFDP1, an E2F1 dimerization partner, inhibits the activating effect of E2F1 and reduces both PITX1 promoter activity and mRNA transcription. Real time RT-PCR results reveal reduced expression of TFDP1 and a similar downregulation of their targets PITX1, BRCA1, CDKN1A, and RAD51 in mid-stage OA chondrocytes. Collectively, our data define a previously uncharacterized role for E2F1 and TFDP1 in the transcriptional regulation of PITX1 in articular chondrocytes. Additional E2F1 targets may be affected in OA pathogenesis.
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            Author and article information

            Journal
            PLoS One
            PLoS ONE
            plos
            plosone
            PLoS ONE
            Public Library of Science (San Francisco, CA USA )
            1932-6203
            23 November 2016
            2016
            : 11
            : 11
            : e0167530
            Article
            PONE-D-16-45647
            10.1371/journal.pone.0167530
            5120844
            27880827
            a955d230-7c99-421b-a4df-e77091ba0561
            © 2016 The PLOS ONE Staff

            This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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