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Raf-1 levels determine the migration rate of primary endometrial stromal cells of patients with endometriosis.

Yotova I, Quan P, Gaba A, Leditznig N, Pateisky P, Kurz C, Tschugguel W - J. Cell. Mol. Med. (2012)

Bottom Line: Raf-1 siRNA knockdown in Co- and Eu-hESC resulted in contraction and decreased migration versus siRNA controls.Lowest Raf-1 levels in Ec-hESC were associated with hyperactivated ROCKII and ezrin/radixin/moesin (E/R/M), impaired migration and a contracted phenotype similar to Raf-1 knockdown in Co- and Eu-hESC.Furthermore, we suggest that in contrast to Co-and Eu-hESC, where the cellular Raf-1 levels regulate the rate of migration, the low cellular Raf-1 content in Ec-hESC, might ensure their restricted migration by preserving the contracted cellular phenotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Medical University of Vienna, Vienna, Austria.

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Related in: MedlinePlus

Western blot analysis of MYPT1 and E/R/M phosphorylation analysed 24 hrs after treatment of hESC with specific Raf-1 inhibitors (ZM336372 and GW5074 – 1 μM) is shown on the left. Representative blots from three independent experiments are given. Graphical representation of pMYPT1 (right panels) is shown as protein levels in% (mean value ± S.D.) relative to their normalized levels (mean value set to 1) in the non-treated controls, *P < 0.05.
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fig05: Western blot analysis of MYPT1 and E/R/M phosphorylation analysed 24 hrs after treatment of hESC with specific Raf-1 inhibitors (ZM336372 and GW5074 – 1 μM) is shown on the left. Representative blots from three independent experiments are given. Graphical representation of pMYPT1 (right panels) is shown as protein levels in% (mean value ± S.D.) relative to their normalized levels (mean value set to 1) in the non-treated controls, *P < 0.05.

Mentions: To further analyse whether Raf-1 activity is required for MYPT1 inactivation, Eu-hESC were treated with Raf-1 kinase inhibitors ZM336372 and GW5074 (1 μM) for 24 hrs. The inhibition of Raf-1 reduced MYPT1 phosphorylation to approximately half (P < 0.05) of the levels of untreated cells (Fig. 5, upper panel), showing that Raf-1 kinase activity is required for the direct regulation of MYPT1 phosphorylation. Similar results were obtained for Co- and Ec-hESC under GW5074 administration (Fig. 5, lower panel).


Raf-1 levels determine the migration rate of primary endometrial stromal cells of patients with endometriosis.

Yotova I, Quan P, Gaba A, Leditznig N, Pateisky P, Kurz C, Tschugguel W - J. Cell. Mol. Med. (2012)

Western blot analysis of MYPT1 and E/R/M phosphorylation analysed 24 hrs after treatment of hESC with specific Raf-1 inhibitors (ZM336372 and GW5074 – 1 μM) is shown on the left. Representative blots from three independent experiments are given. Graphical representation of pMYPT1 (right panels) is shown as protein levels in% (mean value ± S.D.) relative to their normalized levels (mean value set to 1) in the non-treated controls, *P < 0.05.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3822983&req=5

fig05: Western blot analysis of MYPT1 and E/R/M phosphorylation analysed 24 hrs after treatment of hESC with specific Raf-1 inhibitors (ZM336372 and GW5074 – 1 μM) is shown on the left. Representative blots from three independent experiments are given. Graphical representation of pMYPT1 (right panels) is shown as protein levels in% (mean value ± S.D.) relative to their normalized levels (mean value set to 1) in the non-treated controls, *P < 0.05.
Mentions: To further analyse whether Raf-1 activity is required for MYPT1 inactivation, Eu-hESC were treated with Raf-1 kinase inhibitors ZM336372 and GW5074 (1 μM) for 24 hrs. The inhibition of Raf-1 reduced MYPT1 phosphorylation to approximately half (P < 0.05) of the levels of untreated cells (Fig. 5, upper panel), showing that Raf-1 kinase activity is required for the direct regulation of MYPT1 phosphorylation. Similar results were obtained for Co- and Ec-hESC under GW5074 administration (Fig. 5, lower panel).

Bottom Line: Raf-1 siRNA knockdown in Co- and Eu-hESC resulted in contraction and decreased migration versus siRNA controls.Lowest Raf-1 levels in Ec-hESC were associated with hyperactivated ROCKII and ezrin/radixin/moesin (E/R/M), impaired migration and a contracted phenotype similar to Raf-1 knockdown in Co- and Eu-hESC.Furthermore, we suggest that in contrast to Co-and Eu-hESC, where the cellular Raf-1 levels regulate the rate of migration, the low cellular Raf-1 content in Ec-hESC, might ensure their restricted migration by preserving the contracted cellular phenotype.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Medical University of Vienna, Vienna, Austria.

Show MeSH
Related in: MedlinePlus