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The hsa-miR-125a/hsa-let-7e/hsa-miR-99b cluster is potentially implicated in Cystic Fibrosis pathogenesis  Marie-Laure Endale Ahanda, Thierry Bienvenu,

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Presentation on theme: "The hsa-miR-125a/hsa-let-7e/hsa-miR-99b cluster is potentially implicated in Cystic Fibrosis pathogenesis  Marie-Laure Endale Ahanda, Thierry Bienvenu,"— Presentation transcript:

1 The hsa-miR-125a/hsa-let-7e/hsa-miR-99b cluster is potentially implicated in Cystic Fibrosis pathogenesis  Marie-Laure Endale Ahanda, Thierry Bienvenu, Isabelle Sermet-Gaudelus, Laurent Mazzolini, Aleksander Edelman, Rima Zoorob, Noélie Davezac  Journal of Cystic Fibrosis  Volume 14, Issue 5, Pages (September 2015) DOI: /j.jcf Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

2 Fig. 1 DNA sequences of the hsa-miR-125a/hsa-let-7e/hsa-miR-99b cluster in Cystic Fibrosis patients harbouring the DeltaF508-CFTR homozygous mutation. Sequences of pre-hsa-miR-99b, pre-hsa-let-7e and pre-hsa-miR-125a are shown in lower case with sequences of the mature sequence in bold. Three SNPs (boxed) named SNP1 (nt ), SNP2 (nt ) and SNP3 (nt ) are identified in the region. Journal of Cystic Fibrosis  , DOI: ( /j.jcf ) Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

3 Fig. 2 Relative quantification of hsa-miR-125a, hsa-let-7e, and hsa-miR-99b by qRTPCR in pTracer, pTCFWT and pTCFDeltaF508 expressing HeLa cells. Journal of Cystic Fibrosis  , DOI: ( /j.jcf ) Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

4 Fig. 3 Identification of an RNA precursor consisting of hsa-miR-125a, hsa-let-7e and hsa-miR-99b. Journal of Cystic Fibrosis  , DOI: ( /j.jcf ) Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

5 Supplementary Fig. S1 miR-28 and miR-125a are expressed in CFBE and CFPAC and HeLa cells. A and B quantitative RTPCR of miR-28 and miR-125a with CFBE and CFPAC RNA extracts enriched in small RNAs (<200nts). C quantitative RTPCR of miR-28 with the three different HeLa cell lines' total RNA extracts. Journal of Cystic Fibrosis  , DOI: ( /j.jcf ) Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

6 Supplementary Fig. S2 Journal of Cystic Fibrosis  , DOI: ( /j.jcf ) Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions

7 Supplementary Fig. S3 Analysis of Bak, ErbB2, FGFR3 and Sirtuin7 quantities. (A) Immunoblot analysis of Bak and HSP60 was performed on pTracer, pTCFWT and pTCFDeltaF508 HeLa cell extracts. (B) Quantification in arbitrary units was performed with Image J using HSP60 as loading control (n=3). (C) Immunoblots of ErbB2 and actin were realised with pTracer, pTCFWT and pTCFDeltaF508 HeLa cell extracts. (D) Quantification in arbitrary units was performed with Image J using actin as loading control (n=5) *p<0.05 by Student's T test analysis. (E) Immunoblots of FGFR3 and actin were realised with pTRacer, pTCFWT and pTCFDeltaF508 HeLa cell extracts. (F) Quantification in arbitrary units was performed with Image J using actin as loading control (n=7). (G) Immunoblots of Sirtuin7 and actin were realised with pTRacer, pTCFWT and pTCFDeltaF508 HeLa cell extracts. (H) Quantification in arbitrary units was performed with Image J using actin as loading control (n=4). Journal of Cystic Fibrosis  , DOI: ( /j.jcf ) Copyright © 2015 European Cystic Fibrosis Society. Terms and Conditions


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