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Kaisa J. Teittinena, Pauliina Kärkkäinena, Johanna Salonenb, Gunilla Rönnholmc, Hanna Korkeamäkia, Mauno Vihinenb, Nisse Kalkkinenc, Olli Lohia
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],INTRODUCTION.
[object Object],[object Object],[object Object],[object Object],[object Object],types .
[object Object],[object Object],[object Object],[object Object],types .
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Risk factors.
[object Object],[object Object],[object Object],[object Object],Nuclear proteins.
[object Object],[object Object],Nuclear proteins.
[object Object],[object Object],Nucleolar protein-leukemia
[object Object],Objective.
MATERIALES Y METODOS.
[object Object],[object Object],CULTIVO.
[object Object],[object Object],AISLAMIENTO  NUCLEO L OS.
[object Object],[object Object],2 DIGE.
[object Object],[object Object],IDENTIFICACION DE PROTEINAS.
[object Object],[object Object],WESTERN BLOT.
[object Object],[object Object],INMUNOFLUORECENCIA.
RESULTADOS Leucemogénesis nucleolar Fundamento: Aproximación proteómica para buscar proteínas cuya expresión varía entre diferentes tipos de células leucémicas.
[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],RESULTADOS
Niveles de intensidad de cada uno de las  manchas de proteínas, las diferencias en los niveles expresión entre diversas líneas de células leucémicas, se compararon usando de software DeCyderTM. 36    AML/ALL 29    preB/preT RESULTADOS
RESULTADOS Para facilitar el lugar recogiendo los geles 2-DE  fueron post-teñidas con plata. Por último, las manchas de las proteínas seleccionadas fueron digeridas "en gel"  y luego sometidas a nano LC-de masas en tándem análisis de espectrometría
RESULTADOS MASCOTA MS / MS programa de Búsqueda de Ion.
RESULTADOS Expresión nucleolar de PHB Western blot análisis  en los extractos nucleolares de las líneas de células leucémicas.  Igualdad de cargas de las muestras. Expresión nucleolar de TDP-43 y su región terminal N (TDP-43-N) Western blot análisis. Igualdad de cargas de las muestras.
Análisis densiométrico de las intensidades de las bandas en el panel A normalizado a los niveles de CCRF-CEM. RESULTADOS 2.9 y 1.8 a 2.7
Tinción de inmunofluorescencia de PHB en los preparativos nucleolares de CCRF-CEM y FH-CALL-3 células. Tinción de inmunofluorescencia de TDP-43 en los preparativos nucleolar de CCRF-CEM y las células MHH-CALL3. DAPI, 4,6-diamidino-2-phenylindole. RESULTADOS
DISCUSSION Author Affirmation Disagree Agree Ayala YM et al “ Interestingly, both anti-tumorigenic and pro-tumorigenic functions have been reported for PHB, depending on its subcellular localization” X Theiss AL et al “ Loss of TDP-43 in human cells causes genomic instability and increased apoptosis” X Ayala YM et al “ Hyperphosphorylated, ubiquitinated, and cleaved TDP-43 was identified as the main component of cytoplasmic inclusions in two neurodegenerative diseases, frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS)” X Sasaki S et al “ Recently, an immunoelectronmicroscopical analysis revealed that TDP-43 localized specifically to the nucleoli in motor neurons ” X
CONCLUSIONS ,[object Object]
[object Object],CONCLUSIONS
CONCLUSIONS ,[object Object]
CONCLUSIONS ,[object Object]
MAPA CONCEPTUAL
MAPA CONCEPTUAL
MAPA CONCEPTUAL
Mapa conceptual.
Gracias…

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Seminario 2

  • 1. Kaisa J. Teittinena, Pauliina Kärkkäinena, Johanna Salonenb, Gunilla Rönnholmc, Hanna Korkeamäkia, Mauno Vihinenb, Nisse Kalkkinenc, Olli Lohia
  • 2.
  • 3.
  • 4.
  • 5.
  • 6.
  • 7.
  • 8.
  • 9.
  • 11.
  • 12.
  • 13.
  • 14.
  • 15.
  • 16.
  • 17. RESULTADOS Leucemogénesis nucleolar Fundamento: Aproximación proteómica para buscar proteínas cuya expresión varía entre diferentes tipos de células leucémicas.
  • 18.
  • 19. Niveles de intensidad de cada uno de las manchas de proteínas, las diferencias en los niveles expresión entre diversas líneas de células leucémicas, se compararon usando de software DeCyderTM. 36  AML/ALL 29  preB/preT RESULTADOS
  • 20. RESULTADOS Para facilitar el lugar recogiendo los geles 2-DE fueron post-teñidas con plata. Por último, las manchas de las proteínas seleccionadas fueron digeridas "en gel" y luego sometidas a nano LC-de masas en tándem análisis de espectrometría
  • 21. RESULTADOS MASCOTA MS / MS programa de Búsqueda de Ion.
  • 22. RESULTADOS Expresión nucleolar de PHB Western blot análisis en los extractos nucleolares de las líneas de células leucémicas. Igualdad de cargas de las muestras. Expresión nucleolar de TDP-43 y su región terminal N (TDP-43-N) Western blot análisis. Igualdad de cargas de las muestras.
  • 23. Análisis densiométrico de las intensidades de las bandas en el panel A normalizado a los niveles de CCRF-CEM. RESULTADOS 2.9 y 1.8 a 2.7
  • 24. Tinción de inmunofluorescencia de PHB en los preparativos nucleolares de CCRF-CEM y FH-CALL-3 células. Tinción de inmunofluorescencia de TDP-43 en los preparativos nucleolar de CCRF-CEM y las células MHH-CALL3. DAPI, 4,6-diamidino-2-phenylindole. RESULTADOS
  • 25. DISCUSSION Author Affirmation Disagree Agree Ayala YM et al “ Interestingly, both anti-tumorigenic and pro-tumorigenic functions have been reported for PHB, depending on its subcellular localization” X Theiss AL et al “ Loss of TDP-43 in human cells causes genomic instability and increased apoptosis” X Ayala YM et al “ Hyperphosphorylated, ubiquitinated, and cleaved TDP-43 was identified as the main component of cytoplasmic inclusions in two neurodegenerative diseases, frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS)” X Sasaki S et al “ Recently, an immunoelectronmicroscopical analysis revealed that TDP-43 localized specifically to the nucleoli in motor neurons ” X
  • 26.
  • 27.
  • 28.
  • 29.