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Cooperation of distinct Rac-dependent pathways to stabilise E-cadherin adhesion.

Erasmus JC, Welsh NJ, Braga VM - Cell. Signal. (2015)

Bottom Line: In contrast, depletion of another EGFR family member, ErbB3, did not interfere with either process.However, in a strong divergence from EGFR RNAi phenotype, DOCK180 depletion did not perturb actin recruitment or cadherin localisation at junctions.Rather, reduced DOCK180 levels impaired the resistance to mechanical stress of pre-formed cell aggregates.

View Article: PubMed Central - PubMed

Affiliation: Molecular Medicine, National Heart and Lung Institute, Faculty of Medicine, Imperial College London, Sir Alexander Fleming Building, SW7 2AZ London, UK.

No MeSH data available.


EGFR depletion perturbs cell–cell contacts.A, Keratinocytes treated with different siRNA oligos were induced to form cell–cell contacts for 30 min, fixed and stained for E-cadherin. Arrows show E-cadherin at junctions; arrowhead points to perturbed junction.B, Western blot shows knockdown of EGFR or ErbB3 (E3).C, Method for quantification of junction disruption. The length of cell–cell contacts (corner to corner) and the length of E-cadherin staining were obtained for each junction and expressed as a ratio (control junctions = 1).D, Disruption of E-cadherin localization by depletion of EGFR. ErbB3 (E3) and non-targeting oligos (con) were used as controls. N = 3 or N = 2 (ErbB3); about 150 junctions quantified in each replicate. Scale bar = 40 μm. *, p < 0.05.
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f0010: EGFR depletion perturbs cell–cell contacts.A, Keratinocytes treated with different siRNA oligos were induced to form cell–cell contacts for 30 min, fixed and stained for E-cadherin. Arrows show E-cadherin at junctions; arrowhead points to perturbed junction.B, Western blot shows knockdown of EGFR or ErbB3 (E3).C, Method for quantification of junction disruption. The length of cell–cell contacts (corner to corner) and the length of E-cadherin staining were obtained for each junction and expressed as a ratio (control junctions = 1).D, Disruption of E-cadherin localization by depletion of EGFR. ErbB3 (E3) and non-targeting oligos (con) were used as controls. N = 3 or N = 2 (ErbB3); about 150 junctions quantified in each replicate. Scale bar = 40 μm. *, p < 0.05.

Mentions: Strikingly, despite defective Rac activation upon EGFR depletion, junction assembly was not completely abolished (Fig. 2A–B). However, junctions were not uniformly stable: E-cadherin staining was fragmented, not covering the whole extension of each cell–cell contact (Fig. 2A, C, D). In contrast, treatment with ErbB3 or scramble oligos did not impair cadherin localization at junctions (Fig. 2A, D). Thus, Rac signalling downstream of EGFR is necessary for junction maintenance, rather than assembly of new cell–cell contacts.


Cooperation of distinct Rac-dependent pathways to stabilise E-cadherin adhesion.

Erasmus JC, Welsh NJ, Braga VM - Cell. Signal. (2015)

EGFR depletion perturbs cell–cell contacts.A, Keratinocytes treated with different siRNA oligos were induced to form cell–cell contacts for 30 min, fixed and stained for E-cadherin. Arrows show E-cadherin at junctions; arrowhead points to perturbed junction.B, Western blot shows knockdown of EGFR or ErbB3 (E3).C, Method for quantification of junction disruption. The length of cell–cell contacts (corner to corner) and the length of E-cadherin staining were obtained for each junction and expressed as a ratio (control junctions = 1).D, Disruption of E-cadherin localization by depletion of EGFR. ErbB3 (E3) and non-targeting oligos (con) were used as controls. N = 3 or N = 2 (ErbB3); about 150 junctions quantified in each replicate. Scale bar = 40 μm. *, p < 0.05.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4508347&req=5

f0010: EGFR depletion perturbs cell–cell contacts.A, Keratinocytes treated with different siRNA oligos were induced to form cell–cell contacts for 30 min, fixed and stained for E-cadherin. Arrows show E-cadherin at junctions; arrowhead points to perturbed junction.B, Western blot shows knockdown of EGFR or ErbB3 (E3).C, Method for quantification of junction disruption. The length of cell–cell contacts (corner to corner) and the length of E-cadherin staining were obtained for each junction and expressed as a ratio (control junctions = 1).D, Disruption of E-cadherin localization by depletion of EGFR. ErbB3 (E3) and non-targeting oligos (con) were used as controls. N = 3 or N = 2 (ErbB3); about 150 junctions quantified in each replicate. Scale bar = 40 μm. *, p < 0.05.
Mentions: Strikingly, despite defective Rac activation upon EGFR depletion, junction assembly was not completely abolished (Fig. 2A–B). However, junctions were not uniformly stable: E-cadherin staining was fragmented, not covering the whole extension of each cell–cell contact (Fig. 2A, C, D). In contrast, treatment with ErbB3 or scramble oligos did not impair cadherin localization at junctions (Fig. 2A, D). Thus, Rac signalling downstream of EGFR is necessary for junction maintenance, rather than assembly of new cell–cell contacts.

Bottom Line: In contrast, depletion of another EGFR family member, ErbB3, did not interfere with either process.However, in a strong divergence from EGFR RNAi phenotype, DOCK180 depletion did not perturb actin recruitment or cadherin localisation at junctions.Rather, reduced DOCK180 levels impaired the resistance to mechanical stress of pre-formed cell aggregates.

View Article: PubMed Central - PubMed

Affiliation: Molecular Medicine, National Heart and Lung Institute, Faculty of Medicine, Imperial College London, Sir Alexander Fleming Building, SW7 2AZ London, UK.

No MeSH data available.