Structured Review

Millipore filamin a
Disrupting the <t>CaR-filamin</t> interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin
Filamin A, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/filamin a/product/Millipore
Average 93 stars, based on 5 article reviews
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filamin a - by Bioz Stars, 2020-08
93/100 stars

Images

1) Product Images from "The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A"

Article Title: The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A

Journal: The Journal of investigative dermatology

doi: 10.1038/jid.2010.414

Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin
Figure Legend Snippet: Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin

Techniques Used: Activation Assay

Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane
Figure Legend Snippet: Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane

Techniques Used:

Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions
Figure Legend Snippet: Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions

Techniques Used:

2) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

3) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

4) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

5) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

6) Product Images from "HCN1 and HCN2 Proteins Are Expressed in Cochlear Hair Cells"

Article Title: HCN1 and HCN2 Proteins Are Expressed in Cochlear Hair Cells

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M112.375832

Immunoprecipitation by primary antibodies to filamin A, HCN2, and fascin-2. Immunoreactivity for filamin A detected by DAB ( A ) and immunofluorescence ( B ) (mouse MAB1678 raised to human filamin A crossing to rat; 1:1,000, clone PM6/317, Chemicon) was localized to stereocilia of both inner hair cells ( long arrows ) and outer hair cells ( short arrows ) in rat cochlea. Scale bars for A and B, 10 μm. C , lane 1, full-length filamin A (detected with filamin A primary antibody 1:1,000) in immunoprecipitation complex arising from the use of anti-filamin A for immunoprecipitation (1:100) from rat brain lysate. This antibody recognizes unprocessed filamin A (270–280 kDa, arrow , as well as 170-, 150-, and 120-kDa cleavage fragments (Chemicon)). Lane 2, negative control with brain lysate, without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3, standards. D , additional negative control for C . Immunoprecipitation with mouse IgG as negative control probed with antifilamin A. Lane 1, mouse IgG immunoprecipitation of brain lysate + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 2, mouse IgG (no immunoprecipitation) + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 3, denatured mouse IgG electrophoresis, probed with filamin A primary + donkey anti-mouse secondary. No protein was observed corresponding to molecular mass of filamin A ( arrow ). E , Western blot of HCN1 (104 kDa, arrow ) in rat brain lysate (1:50, sc-19706, Santa Cruz Biotechnology), lane 1 ; lane 2 , standards. F , immunoprecipitation by anti-filamin A of full-length HCN1. Lane 1, standards; lane 2 , negative control with brain lysate without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3 , HCN1 immunoprecipitated with anti-filamin A ( arrow , 104 kDa), detected with goat anti-mouse HCN1 (carboxyl terminus) which crosses to rat HCN1 sequence (sc-19706 Santa Cruz Biotechnology). G , complex of filamin A and HCN1 also contained protocadherin 15 CD3. Lane 1, standards; lane 2 , full-length protocadherin 15 CD3 (189 kDa, arrow ) immunoprecipitated with anti-filamin A detected with custom primary antibody (1:10,000) ( arrow ). Goat anti-chick IgY-HRP (Santa Cruz Biotechnology) was used as the secondary antibody (1:10,000). Lane 3, negative control without anti-filamin A immunoprecipitation but with brain lysate and protein A beads and primary and secondary antibodies. H , Western blot of HCN2 in rat brain lysate. Lane 1, standards; lane 2 , HCN2 detected with a rabbit polyclonal antibody (1:200, Alomone). Predicted molecular masses of 95 and 127 kDa corresponding to unglycosylated ( arrow ) and glycosylated HCN2, respectively. I , HCN2 is not detected in the complex immunoprecipitated by anti-filamin A. Lane 1, standards; lane 2 , HCN2 bands observed in Western ( H ) are not detected in immunoprecipitated complex. Bands at 170–180 kDa may correspond to protocadherin 15 CD3, given that 5 of 15 aa in the epitope targeted by the HCN2 antibody are identical in protocadherin 15 CD3, and further given that protocadherin 15 CD3 is highly concentrated in the complex immunoprecipitated by anti-filamin A ( G ). J , anti-HCN2 immunoprecipitates HCN1 from rat brain lysate. Lane 1, standards; lane 2, anti-HCN2 primary antibody (1:33, Alomone) immunoprecipitates HCN1 unglycosylated (104 kDa, arrow ) and glycosylated (120 and 130 kDa) forms detected with anti-HCN1 primary antibody (Santa Cruz Biotechnology); lane 3 , negative control for immunoprecipitation with all components except anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). K , anti-HCN2 immunoprecipitation of HCN1 complex does not include protocadherin 15 CD3. Lane 1, standards; lane 2, anti-protocadherin 15 CD3 primary antibody (1:7,500), goat anti-chick secondary antibody (1:5,000); lane 3 , negative control without anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). No bands were evident for either experimental ( lane 2 ) or negative control ( lane 3 ) with goat anti-chick IgY as the secondary antibody. In a second protocol, a bovine anti-chick secondary antibody (1:5,000) detected an ∼170-kDa protein in both experimental and negative control (not illustrated). L , HCN2 binds to fascin-2 by yeast two-hybrid co-transformation. Row 1, HCN2 carboxyl terminus in bait vector pGBKT7 plus fascin-2 in prey vector pGADT7; row 2 , negative control with fascin-2 in prey construct plus empty bait construct; row 3 , negative control with HCN2 in bait construct plus empty prey construct. The co-transformation screening was performed once, and then the desired colony was re-plated onto another selection media in triplicate, with negative controls. The same concentrations of yeast were plated in the grids for rows 1- 3. M , anti-fascin-2 immunoprecipitation of HCN2. Lane 1, standards; lane 2 , 95 kDa ( arrow ) and 127-kDa bands, corresponding to unglycosylated and glycosylated HCN2, respectively; lane 3 negative control without anti-fascin 2 antibody in immunoprecipitation (brain lysate and beads). N , HCN1 is in fascin-2 immunoprecipitation complex along with HCN2. Lane 1, standards; lane 2 , unglycosylated ( arrow ) and glycosylated forms of HCN1 at 120 kDa. Lane 3, goat IgG immunoprecipitation negative control, with beads and all components except anti-fascin-2 for immunoprecipitation. Three or more immunoprecipitation experiments were carried out for each protein in a given complex.
Figure Legend Snippet: Immunoprecipitation by primary antibodies to filamin A, HCN2, and fascin-2. Immunoreactivity for filamin A detected by DAB ( A ) and immunofluorescence ( B ) (mouse MAB1678 raised to human filamin A crossing to rat; 1:1,000, clone PM6/317, Chemicon) was localized to stereocilia of both inner hair cells ( long arrows ) and outer hair cells ( short arrows ) in rat cochlea. Scale bars for A and B, 10 μm. C , lane 1, full-length filamin A (detected with filamin A primary antibody 1:1,000) in immunoprecipitation complex arising from the use of anti-filamin A for immunoprecipitation (1:100) from rat brain lysate. This antibody recognizes unprocessed filamin A (270–280 kDa, arrow , as well as 170-, 150-, and 120-kDa cleavage fragments (Chemicon)). Lane 2, negative control with brain lysate, without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3, standards. D , additional negative control for C . Immunoprecipitation with mouse IgG as negative control probed with antifilamin A. Lane 1, mouse IgG immunoprecipitation of brain lysate + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 2, mouse IgG (no immunoprecipitation) + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 3, denatured mouse IgG electrophoresis, probed with filamin A primary + donkey anti-mouse secondary. No protein was observed corresponding to molecular mass of filamin A ( arrow ). E , Western blot of HCN1 (104 kDa, arrow ) in rat brain lysate (1:50, sc-19706, Santa Cruz Biotechnology), lane 1 ; lane 2 , standards. F , immunoprecipitation by anti-filamin A of full-length HCN1. Lane 1, standards; lane 2 , negative control with brain lysate without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3 , HCN1 immunoprecipitated with anti-filamin A ( arrow , 104 kDa), detected with goat anti-mouse HCN1 (carboxyl terminus) which crosses to rat HCN1 sequence (sc-19706 Santa Cruz Biotechnology). G , complex of filamin A and HCN1 also contained protocadherin 15 CD3. Lane 1, standards; lane 2 , full-length protocadherin 15 CD3 (189 kDa, arrow ) immunoprecipitated with anti-filamin A detected with custom primary antibody (1:10,000) ( arrow ). Goat anti-chick IgY-HRP (Santa Cruz Biotechnology) was used as the secondary antibody (1:10,000). Lane 3, negative control without anti-filamin A immunoprecipitation but with brain lysate and protein A beads and primary and secondary antibodies. H , Western blot of HCN2 in rat brain lysate. Lane 1, standards; lane 2 , HCN2 detected with a rabbit polyclonal antibody (1:200, Alomone). Predicted molecular masses of 95 and 127 kDa corresponding to unglycosylated ( arrow ) and glycosylated HCN2, respectively. I , HCN2 is not detected in the complex immunoprecipitated by anti-filamin A. Lane 1, standards; lane 2 , HCN2 bands observed in Western ( H ) are not detected in immunoprecipitated complex. Bands at 170–180 kDa may correspond to protocadherin 15 CD3, given that 5 of 15 aa in the epitope targeted by the HCN2 antibody are identical in protocadherin 15 CD3, and further given that protocadherin 15 CD3 is highly concentrated in the complex immunoprecipitated by anti-filamin A ( G ). J , anti-HCN2 immunoprecipitates HCN1 from rat brain lysate. Lane 1, standards; lane 2, anti-HCN2 primary antibody (1:33, Alomone) immunoprecipitates HCN1 unglycosylated (104 kDa, arrow ) and glycosylated (120 and 130 kDa) forms detected with anti-HCN1 primary antibody (Santa Cruz Biotechnology); lane 3 , negative control for immunoprecipitation with all components except anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). K , anti-HCN2 immunoprecipitation of HCN1 complex does not include protocadherin 15 CD3. Lane 1, standards; lane 2, anti-protocadherin 15 CD3 primary antibody (1:7,500), goat anti-chick secondary antibody (1:5,000); lane 3 , negative control without anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). No bands were evident for either experimental ( lane 2 ) or negative control ( lane 3 ) with goat anti-chick IgY as the secondary antibody. In a second protocol, a bovine anti-chick secondary antibody (1:5,000) detected an ∼170-kDa protein in both experimental and negative control (not illustrated). L , HCN2 binds to fascin-2 by yeast two-hybrid co-transformation. Row 1, HCN2 carboxyl terminus in bait vector pGBKT7 plus fascin-2 in prey vector pGADT7; row 2 , negative control with fascin-2 in prey construct plus empty bait construct; row 3 , negative control with HCN2 in bait construct plus empty prey construct. The co-transformation screening was performed once, and then the desired colony was re-plated onto another selection media in triplicate, with negative controls. The same concentrations of yeast were plated in the grids for rows 1- 3. M , anti-fascin-2 immunoprecipitation of HCN2. Lane 1, standards; lane 2 , 95 kDa ( arrow ) and 127-kDa bands, corresponding to unglycosylated and glycosylated HCN2, respectively; lane 3 negative control without anti-fascin 2 antibody in immunoprecipitation (brain lysate and beads). N , HCN1 is in fascin-2 immunoprecipitation complex along with HCN2. Lane 1, standards; lane 2 , unglycosylated ( arrow ) and glycosylated forms of HCN1 at 120 kDa. Lane 3, goat IgG immunoprecipitation negative control, with beads and all components except anti-fascin-2 for immunoprecipitation. Three or more immunoprecipitation experiments were carried out for each protein in a given complex.

Techniques Used: Immunoprecipitation, Immunofluorescence, Negative Control, Electrophoresis, Western Blot, Sequencing, Transformation Assay, Plasmid Preparation, Construct, Selection

7) Product Images from "Reassembly of contractile actin cortex in cell blebs"

Article Title: Reassembly of contractile actin cortex in cell blebs

Journal: The Journal of Cell Biology

doi: 10.1083/jcb.200602085

Actin ultrastructure in retracting blebs. (A) SEM of a blebbing cell with an intact cell membrane. (inset) As the retraction ends, the bleb membrane is crumpled. (B) TEM of the actin cortex of a retracting bleb. The bleb interior is devoid of cytoskeletal structures. Actin is concentrated at the bleb rim and forms a thin shell that is 10–20 nm thick. (C) Actin cytoskeleton of a blebbing cell. (D) Enlargement of boxed area in C. The cortex is formed by an entangled meshwork of actin with a few protuberant knots (arrow). The mesh size is ∼200 nm. Myosin S1 head decoration shows no specific orientation of the actin around the knots. (E) Actin cytoskeleton of a bleb in a dividing HeLa cell. The morphology of blebs is similar to that of filamin-deficient cells, but the mesh is finer. (F) Actin cortex of a rounded HeLa cell. The morphology of the cortex is similar to that of blebs. The entire cell is shown in the inset.
Figure Legend Snippet: Actin ultrastructure in retracting blebs. (A) SEM of a blebbing cell with an intact cell membrane. (inset) As the retraction ends, the bleb membrane is crumpled. (B) TEM of the actin cortex of a retracting bleb. The bleb interior is devoid of cytoskeletal structures. Actin is concentrated at the bleb rim and forms a thin shell that is 10–20 nm thick. (C) Actin cytoskeleton of a blebbing cell. (D) Enlargement of boxed area in C. The cortex is formed by an entangled meshwork of actin with a few protuberant knots (arrow). The mesh size is ∼200 nm. Myosin S1 head decoration shows no specific orientation of the actin around the knots. (E) Actin cytoskeleton of a bleb in a dividing HeLa cell. The morphology of blebs is similar to that of filamin-deficient cells, but the mesh is finer. (F) Actin cortex of a rounded HeLa cell. The morphology of the cortex is similar to that of blebs. The entire cell is shown in the inset.

Techniques Used: Transmission Electron Microscopy

8) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

9) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

10) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

11) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

12) Product Images from "Microaggregate-associated protein involved in invasion of epithelial cells by Mycobacterium avium subsp. hominissuis"

Article Title: Microaggregate-associated protein involved in invasion of epithelial cells by Mycobacterium avium subsp. hominissuis

Journal: Virulence

doi: 10.1080/21505594.2015.1072676

MIP-1 interacts with host protein filamin A. ( A ) Identification of MIP-1 host binding partners by Far-western blot. Lane (1) HEp-1 lysate probed with MIP-1 recombinant protein. Asterisk (*) indicates the position of filamin A on western blot (2) Filamin
Figure Legend Snippet: MIP-1 interacts with host protein filamin A. ( A ) Identification of MIP-1 host binding partners by Far-western blot. Lane (1) HEp-1 lysate probed with MIP-1 recombinant protein. Asterisk (*) indicates the position of filamin A on western blot (2) Filamin

Techniques Used: Binding Assay, Far Western Blot, Recombinant, Western Blot

MIP-1 interacts with host cytoskeletal protein filamin A
Figure Legend Snippet: MIP-1 interacts with host cytoskeletal protein filamin A

Techniques Used:

Microaggregates and planktonic MAH colocalize with host protein filamin A during infection of Hep-2 cells. Immunofluoresnce microscopy was used to examine the colocalization between filamin A and MAC104 microaggregates and planktonic bacteria. White arrows
Figure Legend Snippet: Microaggregates and planktonic MAH colocalize with host protein filamin A during infection of Hep-2 cells. Immunofluoresnce microscopy was used to examine the colocalization between filamin A and MAC104 microaggregates and planktonic bacteria. White arrows

Techniques Used: Infection, Microscopy

13) Product Images from "The Cdc42 Guanine Nucleotide Exchange Factor FGD6 Coordinates Cell Polarity and Endosomal Membrane Recycling in Osteoclasts *"

Article Title: The Cdc42 Guanine Nucleotide Exchange Factor FGD6 Coordinates Cell Polarity and Endosomal Membrane Recycling in Osteoclasts *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M113.504894

FGD6 interacts with IQGAP1, ARHGAP10, Talin-1, and Filamin A at the plasma membrane. A , osteoclasts expressing GFP-FGD6 ( green ) were grown on ODs and stained with DAPI ( blue ) and anti-IQGAP1, anti-ARHGAP10, anti-Talin-1, or anti-Filamin A ( red ). Bars
Figure Legend Snippet: FGD6 interacts with IQGAP1, ARHGAP10, Talin-1, and Filamin A at the plasma membrane. A , osteoclasts expressing GFP-FGD6 ( green ) were grown on ODs and stained with DAPI ( blue ) and anti-IQGAP1, anti-ARHGAP10, anti-Talin-1, or anti-Filamin A ( red ). Bars

Techniques Used: Expressing, Staining

14) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

15) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

16) Product Images from "The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A"

Article Title: The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A

Journal: The Journal of investigative dermatology

doi: 10.1038/jid.2010.414

Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin
Figure Legend Snippet: Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin

Techniques Used: Activation Assay

Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane
Figure Legend Snippet: Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane

Techniques Used:

Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions
Figure Legend Snippet: Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions

Techniques Used:

17) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

18) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

19) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

20) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

21) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

22) Product Images from "Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility"

Article Title: Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility

Journal: Pigment cell & melanoma research

doi: 10.1111/j.1755-148X.2010.00792.x

Klotho negatively regulates the Wnt5A-mediated Filamin A cleavage and invasive potential (A) M93-047 cells were subjected to a matrigel invasion assay. Invasion was measured by counting the cells in the bottom well 48 hrs after seeding on the matrigel-coated transwell filter. For each well, cells were counted in four different fields and averaged. The results are the average of three separate assays in which each condition was performed in triplicate. Error bars are standard error of the mean, * = p
Figure Legend Snippet: Klotho negatively regulates the Wnt5A-mediated Filamin A cleavage and invasive potential (A) M93-047 cells were subjected to a matrigel invasion assay. Invasion was measured by counting the cells in the bottom well 48 hrs after seeding on the matrigel-coated transwell filter. For each well, cells were counted in four different fields and averaged. The results are the average of three separate assays in which each condition was performed in triplicate. Error bars are standard error of the mean, * = p

Techniques Used: Invasion Assay

23) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

24) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

25) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

26) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

27) Product Images from "Novel Automated Tracking Analysis of Particles Subjected to Shear Flow: Kindlin-3 Role in B Cells"

Article Title: Novel Automated Tracking Analysis of Particles Subjected to Shear Flow: Kindlin-3 Role in B Cells

Journal: Biophysical Journal

doi: 10.1016/j.bpj.2013.06.051

The effect of blocking filamin and migfilin function on rolling rates of B cells on E-selectin/ICAM-1. ( A ) Western blot showing expression level of filamins A (reduced to 17.6% ± 3.6, n = 3) and B (reduced to 25.3 ± 2.5%)
Figure Legend Snippet: The effect of blocking filamin and migfilin function on rolling rates of B cells on E-selectin/ICAM-1. ( A ) Western blot showing expression level of filamins A (reduced to 17.6% ± 3.6, n = 3) and B (reduced to 25.3 ± 2.5%)

Techniques Used: Blocking Assay, Western Blot, Expressing

28) Product Images from "HCN1 and HCN2 Proteins Are Expressed in Cochlear Hair Cells"

Article Title: HCN1 and HCN2 Proteins Are Expressed in Cochlear Hair Cells

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M112.375832

Immunoprecipitation by primary antibodies to filamin A, HCN2, and fascin-2. Immunoreactivity for filamin A detected by DAB ( A ) and immunofluorescence ( B ) (mouse MAB1678 raised to human filamin A crossing to rat; 1:1,000, clone PM6/317, Chemicon) was localized to stereocilia of both inner hair cells ( long arrows ) and outer hair cells ( short arrows ) in rat cochlea. Scale bars for A and B, 10 μm. C , lane 1, full-length filamin A (detected with filamin A primary antibody 1:1,000) in immunoprecipitation complex arising from the use of anti-filamin A for immunoprecipitation (1:100) from rat brain lysate. This antibody recognizes unprocessed filamin A (270–280 kDa, arrow , as well as 170-, 150-, and 120-kDa cleavage fragments (Chemicon)). Lane 2, negative control with brain lysate, without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3, standards. D , additional negative control for C . Immunoprecipitation with mouse IgG as negative control probed with antifilamin A. Lane 1, mouse IgG immunoprecipitation of brain lysate + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 2, mouse IgG (no immunoprecipitation) + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 3, denatured mouse IgG electrophoresis, probed with filamin A primary + donkey anti-mouse secondary. No protein was observed corresponding to molecular mass of filamin A ( arrow ). E , Western blot of HCN1 (104 kDa, arrow ) in rat brain lysate (1:50, sc-19706, Santa Cruz Biotechnology), lane 1 ; lane 2 , standards. F , immunoprecipitation by anti-filamin A of full-length HCN1. Lane 1, standards; lane 2 , negative control with brain lysate without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3 , HCN1 immunoprecipitated with anti-filamin A ( arrow , 104 kDa), detected with goat anti-mouse HCN1 (carboxyl terminus) which crosses to rat HCN1 sequence (sc-19706 Santa Cruz Biotechnology). G , complex of filamin A and HCN1 also contained protocadherin 15 CD3. Lane 1, standards; lane 2 , full-length protocadherin 15 CD3 (189 kDa, arrow ) immunoprecipitated with anti-filamin A detected with custom primary antibody (1:10,000) ( arrow ). Goat anti-chick IgY-HRP (Santa Cruz Biotechnology) was used as the secondary antibody (1:10,000). Lane 3, negative control without anti-filamin A immunoprecipitation but with brain lysate and protein A beads and primary and secondary antibodies. H , Western blot of HCN2 in rat brain lysate. Lane 1, standards; lane 2 , HCN2 detected with a rabbit polyclonal antibody (1:200, Alomone). Predicted molecular masses of 95 and 127 kDa corresponding to unglycosylated ( arrow ) and glycosylated HCN2, respectively. I , HCN2 is not detected in the complex immunoprecipitated by anti-filamin A. Lane 1, standards; lane 2 , HCN2 bands observed in Western ( H ) are not detected in immunoprecipitated complex. Bands at 170–180 kDa may correspond to protocadherin 15 CD3, given that 5 of 15 aa in the epitope targeted by the HCN2 antibody are identical in protocadherin 15 CD3, and further given that protocadherin 15 CD3 is highly concentrated in the complex immunoprecipitated by anti-filamin A ( G ). J , anti-HCN2 immunoprecipitates HCN1 from rat brain lysate. Lane 1, standards; lane 2, anti-HCN2 primary antibody (1:33, Alomone) immunoprecipitates HCN1 unglycosylated (104 kDa, arrow ) and glycosylated (120 and 130 kDa) forms detected with anti-HCN1 primary antibody (Santa Cruz Biotechnology); lane 3 , negative control for immunoprecipitation with all components except anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). K , anti-HCN2 immunoprecipitation of HCN1 complex does not include protocadherin 15 CD3. Lane 1, standards; lane 2, anti-protocadherin 15 CD3 primary antibody (1:7,500), goat anti-chick secondary antibody (1:5,000); lane 3 , negative control without anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). No bands were evident for either experimental ( lane 2 ) or negative control ( lane 3 ) with goat anti-chick IgY as the secondary antibody. In a second protocol, a bovine anti-chick secondary antibody (1:5,000) detected an ∼170-kDa protein in both experimental and negative control (not illustrated). L , HCN2 binds to fascin-2 by yeast two-hybrid co-transformation. Row 1, HCN2 carboxyl terminus in bait vector pGBKT7 plus fascin-2 in prey vector pGADT7; row 2 , negative control with fascin-2 in prey construct plus empty bait construct; row 3 , negative control with HCN2 in bait construct plus empty prey construct. The co-transformation screening was performed once, and then the desired colony was re-plated onto another selection media in triplicate, with negative controls. The same concentrations of yeast were plated in the grids for rows 1- 3. M , anti-fascin-2 immunoprecipitation of HCN2. Lane 1, standards; lane 2 , 95 kDa ( arrow ) and 127-kDa bands, corresponding to unglycosylated and glycosylated HCN2, respectively; lane 3 negative control without anti-fascin 2 antibody in immunoprecipitation (brain lysate and beads). N , HCN1 is in fascin-2 immunoprecipitation complex along with HCN2. Lane 1, standards; lane 2 , unglycosylated ( arrow ) and glycosylated forms of HCN1 at 120 kDa. Lane 3, goat IgG immunoprecipitation negative control, with beads and all components except anti-fascin-2 for immunoprecipitation. Three or more immunoprecipitation experiments were carried out for each protein in a given complex.
Figure Legend Snippet: Immunoprecipitation by primary antibodies to filamin A, HCN2, and fascin-2. Immunoreactivity for filamin A detected by DAB ( A ) and immunofluorescence ( B ) (mouse MAB1678 raised to human filamin A crossing to rat; 1:1,000, clone PM6/317, Chemicon) was localized to stereocilia of both inner hair cells ( long arrows ) and outer hair cells ( short arrows ) in rat cochlea. Scale bars for A and B, 10 μm. C , lane 1, full-length filamin A (detected with filamin A primary antibody 1:1,000) in immunoprecipitation complex arising from the use of anti-filamin A for immunoprecipitation (1:100) from rat brain lysate. This antibody recognizes unprocessed filamin A (270–280 kDa, arrow , as well as 170-, 150-, and 120-kDa cleavage fragments (Chemicon)). Lane 2, negative control with brain lysate, without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3, standards. D , additional negative control for C . Immunoprecipitation with mouse IgG as negative control probed with antifilamin A. Lane 1, mouse IgG immunoprecipitation of brain lysate + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 2, mouse IgG (no immunoprecipitation) + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 3, denatured mouse IgG electrophoresis, probed with filamin A primary + donkey anti-mouse secondary. No protein was observed corresponding to molecular mass of filamin A ( arrow ). E , Western blot of HCN1 (104 kDa, arrow ) in rat brain lysate (1:50, sc-19706, Santa Cruz Biotechnology), lane 1 ; lane 2 , standards. F , immunoprecipitation by anti-filamin A of full-length HCN1. Lane 1, standards; lane 2 , negative control with brain lysate without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3 , HCN1 immunoprecipitated with anti-filamin A ( arrow , 104 kDa), detected with goat anti-mouse HCN1 (carboxyl terminus) which crosses to rat HCN1 sequence (sc-19706 Santa Cruz Biotechnology). G , complex of filamin A and HCN1 also contained protocadherin 15 CD3. Lane 1, standards; lane 2 , full-length protocadherin 15 CD3 (189 kDa, arrow ) immunoprecipitated with anti-filamin A detected with custom primary antibody (1:10,000) ( arrow ). Goat anti-chick IgY-HRP (Santa Cruz Biotechnology) was used as the secondary antibody (1:10,000). Lane 3, negative control without anti-filamin A immunoprecipitation but with brain lysate and protein A beads and primary and secondary antibodies. H , Western blot of HCN2 in rat brain lysate. Lane 1, standards; lane 2 , HCN2 detected with a rabbit polyclonal antibody (1:200, Alomone). Predicted molecular masses of 95 and 127 kDa corresponding to unglycosylated ( arrow ) and glycosylated HCN2, respectively. I , HCN2 is not detected in the complex immunoprecipitated by anti-filamin A. Lane 1, standards; lane 2 , HCN2 bands observed in Western ( H ) are not detected in immunoprecipitated complex. Bands at 170–180 kDa may correspond to protocadherin 15 CD3, given that 5 of 15 aa in the epitope targeted by the HCN2 antibody are identical in protocadherin 15 CD3, and further given that protocadherin 15 CD3 is highly concentrated in the complex immunoprecipitated by anti-filamin A ( G ). J , anti-HCN2 immunoprecipitates HCN1 from rat brain lysate. Lane 1, standards; lane 2, anti-HCN2 primary antibody (1:33, Alomone) immunoprecipitates HCN1 unglycosylated (104 kDa, arrow ) and glycosylated (120 and 130 kDa) forms detected with anti-HCN1 primary antibody (Santa Cruz Biotechnology); lane 3 , negative control for immunoprecipitation with all components except anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). K , anti-HCN2 immunoprecipitation of HCN1 complex does not include protocadherin 15 CD3. Lane 1, standards; lane 2, anti-protocadherin 15 CD3 primary antibody (1:7,500), goat anti-chick secondary antibody (1:5,000); lane 3 , negative control without anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). No bands were evident for either experimental ( lane 2 ) or negative control ( lane 3 ) with goat anti-chick IgY as the secondary antibody. In a second protocol, a bovine anti-chick secondary antibody (1:5,000) detected an ∼170-kDa protein in both experimental and negative control (not illustrated). L , HCN2 binds to fascin-2 by yeast two-hybrid co-transformation. Row 1, HCN2 carboxyl terminus in bait vector pGBKT7 plus fascin-2 in prey vector pGADT7; row 2 , negative control with fascin-2 in prey construct plus empty bait construct; row 3 , negative control with HCN2 in bait construct plus empty prey construct. The co-transformation screening was performed once, and then the desired colony was re-plated onto another selection media in triplicate, with negative controls. The same concentrations of yeast were plated in the grids for rows 1- 3. M , anti-fascin-2 immunoprecipitation of HCN2. Lane 1, standards; lane 2 , 95 kDa ( arrow ) and 127-kDa bands, corresponding to unglycosylated and glycosylated HCN2, respectively; lane 3 negative control without anti-fascin 2 antibody in immunoprecipitation (brain lysate and beads). N , HCN1 is in fascin-2 immunoprecipitation complex along with HCN2. Lane 1, standards; lane 2 , unglycosylated ( arrow ) and glycosylated forms of HCN1 at 120 kDa. Lane 3, goat IgG immunoprecipitation negative control, with beads and all components except anti-fascin-2 for immunoprecipitation. Three or more immunoprecipitation experiments were carried out for each protein in a given complex.

Techniques Used: Immunoprecipitation, Immunofluorescence, Negative Control, Electrophoresis, Western Blot, Sequencing, Transformation Assay, Plasmid Preparation, Construct, Selection

29) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

30) Product Images from "Filamin B Plays a Key Role in Vascular Endothelial Growth Factor-induced Endothelial Cell Motility through Its Interaction with Rac-1 and Vav-2 *"

Article Title: Filamin B Plays a Key Role in Vascular Endothelial Growth Factor-induced Endothelial Cell Motility through Its Interaction with Rac-1 and Vav-2 *

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M109.062984

Filamin A- and B-specific knockdown by siRNA transfection in endothelial cells. A , Northern blotting analysis. HUVEC, HT29, and HEK-293 (human embryonic kidney) cells are shown. Filamin B, filamin A, and glyceraldehyde-3-phosphate dehydrogenase (as a
Figure Legend Snippet: Filamin A- and B-specific knockdown by siRNA transfection in endothelial cells. A , Northern blotting analysis. HUVEC, HT29, and HEK-293 (human embryonic kidney) cells are shown. Filamin B, filamin A, and glyceraldehyde-3-phosphate dehydrogenase (as a

Techniques Used: Transfection, Northern Blot

Filamin B interacts with Rac-1 in basal conditions and with Rac-1, VEGFR2, and integrin αvβ5 in VEGF-stimulated cells. A , cells in basal conditions ( Basal ) or stimulated with 20 ng/ml VEGF for 5 min (+ VEGF ) were lysed, and VEGFR2, Rac-1,
Figure Legend Snippet: Filamin B interacts with Rac-1 in basal conditions and with Rac-1, VEGFR2, and integrin αvβ5 in VEGF-stimulated cells. A , cells in basal conditions ( Basal ) or stimulated with 20 ng/ml VEGF for 5 min (+ VEGF ) were lysed, and VEGFR2, Rac-1,

Techniques Used:

Filamin B depletion causes overstimulation of Rac-1 and PAK-4/5/6 basal activities in the absence of any effect on ERK1/2. A , Rac-1 activity was measured using a G-LISA assay. Unrelated siRNA ( UNr )- or siRNA-FLNB3-transfected cell lysates were incubated
Figure Legend Snippet: Filamin B depletion causes overstimulation of Rac-1 and PAK-4/5/6 basal activities in the absence of any effect on ERK1/2. A , Rac-1 activity was measured using a G-LISA assay. Unrelated siRNA ( UNr )- or siRNA-FLNB3-transfected cell lysates were incubated

Techniques Used: Activity Assay, Transfection, Incubation

Filamin B is essential for VEGF-induced in vitro angiogenesis and endothelial cell migration. A , HUVEC were transiently transfected with an unrelated control siRNA, siRNA-FLNA1 and -FLNA2, or siRNA-FLNB2 and -FLNB3, as described. After 24 h, cells were
Figure Legend Snippet: Filamin B is essential for VEGF-induced in vitro angiogenesis and endothelial cell migration. A , HUVEC were transiently transfected with an unrelated control siRNA, siRNA-FLNA1 and -FLNA2, or siRNA-FLNB2 and -FLNB3, as described. After 24 h, cells were

Techniques Used: In Vitro, Migration, Transfection

Filamin B binds Vav-2 in basal conditions and after VEGF stimulation, and its depletion abolishes Vav-2 phosphorylation in the absence of any effect of Src phosphorylation. A , HUVEC were starved for 16 h and then stimulated or not with 10 ng/ml VEGF for
Figure Legend Snippet: Filamin B binds Vav-2 in basal conditions and after VEGF stimulation, and its depletion abolishes Vav-2 phosphorylation in the absence of any effect of Src phosphorylation. A , HUVEC were starved for 16 h and then stimulated or not with 10 ng/ml VEGF for

Techniques Used:

Filamin B depletion blocks Rac-1 VEGF-induced translocation to plasma membrane. A , HUVEC were transiently transfected with an unrelated control siRNA ( Unr ) or siRNA-FLNB3 as described, in the absence or presence of 20 ng/ml VEGF for 30 min. Cells were
Figure Legend Snippet: Filamin B depletion blocks Rac-1 VEGF-induced translocation to plasma membrane. A , HUVEC were transiently transfected with an unrelated control siRNA ( Unr ) or siRNA-FLNB3 as described, in the absence or presence of 20 ng/ml VEGF for 30 min. Cells were

Techniques Used: Translocation Assay, Transfection

Filamin B knockdown leads to an increase in focal adhesions. A , immunolocalization of F-actin ( blue ) and filamin A ( red ) in HUVEC transiently transfected with an unrelated control siRNA ( Unr ) or siRNA-FLNB3 as described, in the absence (− VEGF
Figure Legend Snippet: Filamin B knockdown leads to an increase in focal adhesions. A , immunolocalization of F-actin ( blue ) and filamin A ( red ) in HUVEC transiently transfected with an unrelated control siRNA ( Unr ) or siRNA-FLNB3 as described, in the absence (− VEGF

Techniques Used: Transfection

31) Product Images from "Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility"

Article Title: Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility

Journal: Pigment cell & melanoma research

doi: 10.1111/j.1755-148X.2010.00792.x

Klotho negatively regulates the Wnt5A-mediated Filamin A cleavage and invasive potential (A) M93-047 cells were subjected to a matrigel invasion assay. Invasion was measured by counting the cells in the bottom well 48 hrs after seeding on the matrigel-coated transwell filter. For each well, cells were counted in four different fields and averaged. The results are the average of three separate assays in which each condition was performed in triplicate. Error bars are standard error of the mean, * = p
Figure Legend Snippet: Klotho negatively regulates the Wnt5A-mediated Filamin A cleavage and invasive potential (A) M93-047 cells were subjected to a matrigel invasion assay. Invasion was measured by counting the cells in the bottom well 48 hrs after seeding on the matrigel-coated transwell filter. For each well, cells were counted in four different fields and averaged. The results are the average of three separate assays in which each condition was performed in triplicate. Error bars are standard error of the mean, * = p

Techniques Used: Invasion Assay

32) Product Images from "Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility"

Article Title: Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility

Journal: Pigment cell & melanoma research

doi: 10.1111/j.1755-148X.2010.00792.x

Klotho negatively regulates the Wnt5A-mediated Filamin A cleavage and invasive potential (A) M93-047 cells were subjected to a matrigel invasion assay. Invasion was measured by counting the cells in the bottom well 48 hrs after seeding on the matrigel-coated transwell filter. For each well, cells were counted in four different fields and averaged. The results are the average of three separate assays in which each condition was performed in triplicate. Error bars are standard error of the mean, * = p
Figure Legend Snippet: Klotho negatively regulates the Wnt5A-mediated Filamin A cleavage and invasive potential (A) M93-047 cells were subjected to a matrigel invasion assay. Invasion was measured by counting the cells in the bottom well 48 hrs after seeding on the matrigel-coated transwell filter. For each well, cells were counted in four different fields and averaged. The results are the average of three separate assays in which each condition was performed in triplicate. Error bars are standard error of the mean, * = p

Techniques Used: Invasion Assay

33) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

34) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

35) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

36) Product Images from "Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes"

Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

Journal: Journal of dermatological science

doi: 10.1016/j.jdermsci.2013.09.007

Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control
Figure Legend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

Techniques Used: Inhibition, Expressing, Activation Assay, Transfection

Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)
Figure Legend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

Techniques Used: Expressing, Activation Assay, Infection

Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

Techniques Used: Inhibition, Expressing, Transfection

Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies
Figure Legend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

Techniques Used: Fluorescence, Immunostaining

Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors
Figure Legend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

Techniques Used:

Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin
Figure Legend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

Techniques Used: Inhibition, Expressing, Transfection, Immunoprecipitation

Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected
Figure Legend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

Techniques Used: Infection

37) Product Images from "HCN1 and HCN2 Proteins Are Expressed in Cochlear Hair Cells"

Article Title: HCN1 and HCN2 Proteins Are Expressed in Cochlear Hair Cells

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M112.375832

Immunoprecipitation by primary antibodies to filamin A, HCN2, and fascin-2. Immunoreactivity for filamin A detected by DAB ( A ) and immunofluorescence ( B ) (mouse MAB1678 raised to human filamin A crossing to rat; 1:1,000, clone PM6/317, Chemicon) was localized to stereocilia of both inner hair cells ( long arrows ) and outer hair cells ( short arrows ) in rat cochlea. Scale bars for A and B, 10 μm. C , lane 1, full-length filamin A (detected with filamin A primary antibody 1:1,000) in immunoprecipitation complex arising from the use of anti-filamin A for immunoprecipitation (1:100) from rat brain lysate. This antibody recognizes unprocessed filamin A (270–280 kDa, arrow , as well as 170-, 150-, and 120-kDa cleavage fragments (Chemicon)). Lane 2, negative control with brain lysate, without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3, standards. D , additional negative control for C . Immunoprecipitation with mouse IgG as negative control probed with antifilamin A. Lane 1, mouse IgG immunoprecipitation of brain lysate + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 2, mouse IgG (no immunoprecipitation) + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 3, denatured mouse IgG electrophoresis, probed with filamin A primary + donkey anti-mouse secondary. No protein was observed corresponding to molecular mass of filamin A ( arrow ). E , Western blot of HCN1 (104 kDa, arrow ) in rat brain lysate (1:50, sc-19706, Santa Cruz Biotechnology), lane 1 ; lane 2 , standards. F , immunoprecipitation by anti-filamin A of full-length HCN1. Lane 1, standards; lane 2 , negative control with brain lysate without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3 , HCN1 immunoprecipitated with anti-filamin A ( arrow , 104 kDa), detected with goat anti-mouse HCN1 (carboxyl terminus) which crosses to rat HCN1 sequence (sc-19706 Santa Cruz Biotechnology). G , complex of filamin A and HCN1 also contained protocadherin 15 CD3. Lane 1, standards; lane 2 , full-length protocadherin 15 CD3 (189 kDa, arrow ) immunoprecipitated with anti-filamin A detected with custom primary antibody (1:10,000) ( arrow ). Goat anti-chick IgY-HRP (Santa Cruz Biotechnology) was used as the secondary antibody (1:10,000). Lane 3, negative control without anti-filamin A immunoprecipitation but with brain lysate and protein A beads and primary and secondary antibodies. H , Western blot of HCN2 in rat brain lysate. Lane 1, standards; lane 2 , HCN2 detected with a rabbit polyclonal antibody (1:200, Alomone). Predicted molecular masses of 95 and 127 kDa corresponding to unglycosylated ( arrow ) and glycosylated HCN2, respectively. I , HCN2 is not detected in the complex immunoprecipitated by anti-filamin A. Lane 1, standards; lane 2 , HCN2 bands observed in Western ( H ) are not detected in immunoprecipitated complex. Bands at 170–180 kDa may correspond to protocadherin 15 CD3, given that 5 of 15 aa in the epitope targeted by the HCN2 antibody are identical in protocadherin 15 CD3, and further given that protocadherin 15 CD3 is highly concentrated in the complex immunoprecipitated by anti-filamin A ( G ). J , anti-HCN2 immunoprecipitates HCN1 from rat brain lysate. Lane 1, standards; lane 2, anti-HCN2 primary antibody (1:33, Alomone) immunoprecipitates HCN1 unglycosylated (104 kDa, arrow ) and glycosylated (120 and 130 kDa) forms detected with anti-HCN1 primary antibody (Santa Cruz Biotechnology); lane 3 , negative control for immunoprecipitation with all components except anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). K , anti-HCN2 immunoprecipitation of HCN1 complex does not include protocadherin 15 CD3. Lane 1, standards; lane 2, anti-protocadherin 15 CD3 primary antibody (1:7,500), goat anti-chick secondary antibody (1:5,000); lane 3 , negative control without anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). No bands were evident for either experimental ( lane 2 ) or negative control ( lane 3 ) with goat anti-chick IgY as the secondary antibody. In a second protocol, a bovine anti-chick secondary antibody (1:5,000) detected an ∼170-kDa protein in both experimental and negative control (not illustrated). L , HCN2 binds to fascin-2 by yeast two-hybrid co-transformation. Row 1, HCN2 carboxyl terminus in bait vector pGBKT7 plus fascin-2 in prey vector pGADT7; row 2 , negative control with fascin-2 in prey construct plus empty bait construct; row 3 , negative control with HCN2 in bait construct plus empty prey construct. The co-transformation screening was performed once, and then the desired colony was re-plated onto another selection media in triplicate, with negative controls. The same concentrations of yeast were plated in the grids for rows 1- 3. M , anti-fascin-2 immunoprecipitation of HCN2. Lane 1, standards; lane 2 , 95 kDa ( arrow ) and 127-kDa bands, corresponding to unglycosylated and glycosylated HCN2, respectively; lane 3 negative control without anti-fascin 2 antibody in immunoprecipitation (brain lysate and beads). N , HCN1 is in fascin-2 immunoprecipitation complex along with HCN2. Lane 1, standards; lane 2 , unglycosylated ( arrow ) and glycosylated forms of HCN1 at 120 kDa. Lane 3, goat IgG immunoprecipitation negative control, with beads and all components except anti-fascin-2 for immunoprecipitation. Three or more immunoprecipitation experiments were carried out for each protein in a given complex.
Figure Legend Snippet: Immunoprecipitation by primary antibodies to filamin A, HCN2, and fascin-2. Immunoreactivity for filamin A detected by DAB ( A ) and immunofluorescence ( B ) (mouse MAB1678 raised to human filamin A crossing to rat; 1:1,000, clone PM6/317, Chemicon) was localized to stereocilia of both inner hair cells ( long arrows ) and outer hair cells ( short arrows ) in rat cochlea. Scale bars for A and B, 10 μm. C , lane 1, full-length filamin A (detected with filamin A primary antibody 1:1,000) in immunoprecipitation complex arising from the use of anti-filamin A for immunoprecipitation (1:100) from rat brain lysate. This antibody recognizes unprocessed filamin A (270–280 kDa, arrow , as well as 170-, 150-, and 120-kDa cleavage fragments (Chemicon)). Lane 2, negative control with brain lysate, without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3, standards. D , additional negative control for C . Immunoprecipitation with mouse IgG as negative control probed with antifilamin A. Lane 1, mouse IgG immunoprecipitation of brain lysate + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 2, mouse IgG (no immunoprecipitation) + beads, probed with filamin A primary + donkey anti-mouse secondary. Lane 3, denatured mouse IgG electrophoresis, probed with filamin A primary + donkey anti-mouse secondary. No protein was observed corresponding to molecular mass of filamin A ( arrow ). E , Western blot of HCN1 (104 kDa, arrow ) in rat brain lysate (1:50, sc-19706, Santa Cruz Biotechnology), lane 1 ; lane 2 , standards. F , immunoprecipitation by anti-filamin A of full-length HCN1. Lane 1, standards; lane 2 , negative control with brain lysate without anti-filamin A immunoprecipitation + beads + primary and secondary antibodies; lane 3 , HCN1 immunoprecipitated with anti-filamin A ( arrow , 104 kDa), detected with goat anti-mouse HCN1 (carboxyl terminus) which crosses to rat HCN1 sequence (sc-19706 Santa Cruz Biotechnology). G , complex of filamin A and HCN1 also contained protocadherin 15 CD3. Lane 1, standards; lane 2 , full-length protocadherin 15 CD3 (189 kDa, arrow ) immunoprecipitated with anti-filamin A detected with custom primary antibody (1:10,000) ( arrow ). Goat anti-chick IgY-HRP (Santa Cruz Biotechnology) was used as the secondary antibody (1:10,000). Lane 3, negative control without anti-filamin A immunoprecipitation but with brain lysate and protein A beads and primary and secondary antibodies. H , Western blot of HCN2 in rat brain lysate. Lane 1, standards; lane 2 , HCN2 detected with a rabbit polyclonal antibody (1:200, Alomone). Predicted molecular masses of 95 and 127 kDa corresponding to unglycosylated ( arrow ) and glycosylated HCN2, respectively. I , HCN2 is not detected in the complex immunoprecipitated by anti-filamin A. Lane 1, standards; lane 2 , HCN2 bands observed in Western ( H ) are not detected in immunoprecipitated complex. Bands at 170–180 kDa may correspond to protocadherin 15 CD3, given that 5 of 15 aa in the epitope targeted by the HCN2 antibody are identical in protocadherin 15 CD3, and further given that protocadherin 15 CD3 is highly concentrated in the complex immunoprecipitated by anti-filamin A ( G ). J , anti-HCN2 immunoprecipitates HCN1 from rat brain lysate. Lane 1, standards; lane 2, anti-HCN2 primary antibody (1:33, Alomone) immunoprecipitates HCN1 unglycosylated (104 kDa, arrow ) and glycosylated (120 and 130 kDa) forms detected with anti-HCN1 primary antibody (Santa Cruz Biotechnology); lane 3 , negative control for immunoprecipitation with all components except anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). K , anti-HCN2 immunoprecipitation of HCN1 complex does not include protocadherin 15 CD3. Lane 1, standards; lane 2, anti-protocadherin 15 CD3 primary antibody (1:7,500), goat anti-chick secondary antibody (1:5,000); lane 3 , negative control without anti-HCN2 antibody for immunoprecipitation (brain lysate and beads). No bands were evident for either experimental ( lane 2 ) or negative control ( lane 3 ) with goat anti-chick IgY as the secondary antibody. In a second protocol, a bovine anti-chick secondary antibody (1:5,000) detected an ∼170-kDa protein in both experimental and negative control (not illustrated). L , HCN2 binds to fascin-2 by yeast two-hybrid co-transformation. Row 1, HCN2 carboxyl terminus in bait vector pGBKT7 plus fascin-2 in prey vector pGADT7; row 2 , negative control with fascin-2 in prey construct plus empty bait construct; row 3 , negative control with HCN2 in bait construct plus empty prey construct. The co-transformation screening was performed once, and then the desired colony was re-plated onto another selection media in triplicate, with negative controls. The same concentrations of yeast were plated in the grids for rows 1- 3. M , anti-fascin-2 immunoprecipitation of HCN2. Lane 1, standards; lane 2 , 95 kDa ( arrow ) and 127-kDa bands, corresponding to unglycosylated and glycosylated HCN2, respectively; lane 3 negative control without anti-fascin 2 antibody in immunoprecipitation (brain lysate and beads). N , HCN1 is in fascin-2 immunoprecipitation complex along with HCN2. Lane 1, standards; lane 2 , unglycosylated ( arrow ) and glycosylated forms of HCN1 at 120 kDa. Lane 3, goat IgG immunoprecipitation negative control, with beads and all components except anti-fascin-2 for immunoprecipitation. Three or more immunoprecipitation experiments were carried out for each protein in a given complex.

Techniques Used: Immunoprecipitation, Immunofluorescence, Negative Control, Electrophoresis, Western Blot, Sequencing, Transformation Assay, Plasmid Preparation, Construct, Selection

38) Product Images from "The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A"

Article Title: The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A

Journal: The Journal of investigative dermatology

doi: 10.1038/jid.2010.414

Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin
Figure Legend Snippet: Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin

Techniques Used: Activation Assay

Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane
Figure Legend Snippet: Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane

Techniques Used:

Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions
Figure Legend Snippet: Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions

Techniques Used:

39) Product Images from "The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A"

Article Title: The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A

Journal: The Journal of investigative dermatology

doi: 10.1038/jid.2010.414

Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin
Figure Legend Snippet: Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin

Techniques Used: Activation Assay

Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane
Figure Legend Snippet: Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane

Techniques Used:

Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions
Figure Legend Snippet: Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions

Techniques Used:

40) Product Images from "The Cytoskeleton Protein Filamin-A is Required for an Efficient Recombinational DNA Double Strand Break Repair"

Article Title: The Cytoskeleton Protein Filamin-A is Required for an Efficient Recombinational DNA Double Strand Break Repair

Journal: Cancer research

doi: 10.1158/0008-5472.CAN-09-2177

Filamin-A deficiency reduces RAD51 recruitment to DNA damage. Four hours after 2, 4, or 8 Gy of γ-irradiation, the RAD51 nuclear foci and chromatin-bound RAD51 protein were measured. Panel A is a western blot showing that lack of filamin-A in M2 cells does not alter the total RAD51 protein levels either before or after the irradiation. Panel B shows the average numbers of RAD51 nuclear foci per cells. Panel C shows the percentage of cells with five or more RAD51 foci. A7 and M2 cells in exponential phase were grown on cover slips and irradiated with 0, 2, 4 or 8 Gy of gamma radiation. Cells were fixed 4 hours after irradiation and fluorescent immunostaining was performed with anti-RAD51 antibody. RAD51 focus number in individual cells was scored under a fluorescent microscope. At least 250 cells in each experiment were analyzed and data shown are averages of 3 independent experiments. The p-values indicate statistic significance based on t -tests. Panel D shows the chromatin-bound RAD51 proteins at time points after irradiation. Filamin-A deficient cells have less chromatin-bound RAD51 protein between 4–12 hours after irradiation. Histone-2A (H2A) was used as a loading control for chromatin-bound proteins.
Figure Legend Snippet: Filamin-A deficiency reduces RAD51 recruitment to DNA damage. Four hours after 2, 4, or 8 Gy of γ-irradiation, the RAD51 nuclear foci and chromatin-bound RAD51 protein were measured. Panel A is a western blot showing that lack of filamin-A in M2 cells does not alter the total RAD51 protein levels either before or after the irradiation. Panel B shows the average numbers of RAD51 nuclear foci per cells. Panel C shows the percentage of cells with five or more RAD51 foci. A7 and M2 cells in exponential phase were grown on cover slips and irradiated with 0, 2, 4 or 8 Gy of gamma radiation. Cells were fixed 4 hours after irradiation and fluorescent immunostaining was performed with anti-RAD51 antibody. RAD51 focus number in individual cells was scored under a fluorescent microscope. At least 250 cells in each experiment were analyzed and data shown are averages of 3 independent experiments. The p-values indicate statistic significance based on t -tests. Panel D shows the chromatin-bound RAD51 proteins at time points after irradiation. Filamin-A deficient cells have less chromatin-bound RAD51 protein between 4–12 hours after irradiation. Histone-2A (H2A) was used as a loading control for chromatin-bound proteins.

Techniques Used: Irradiation, Western Blot, Immunostaining, Microscopy

Filamin-A deficiency impairs the DNA double strand break repair. Panel A shows the numbers of γH2AX foci per cell in A7 and M2 cells. At time points after the cells were treated with γ-radiation (panel A), cells were fixed and stained by immunoflurescent techniques with an anti-γH2AX antibody. The number of γH2AX foci was counted in > 400 cells per time point in each experiment. Data shown are averages and standard errors of three experiments. Panel B shows the total γH2AX protein level measured by anti-γH2AX western blot at time points after irradiation (4 Gy) in C8161 and MB231 cells. The dose and times after irradiation are shown on the top of the panel. “FLNa-shRNA” indicates the whether the cells express filamin-A-specific shRNA. Beta-actin was used as a loading control. Panel C shows the total number of unstable chromosome aberrations per metaphase cell in A7 and M2. At 15 min, 8 hours, and 24 hours after 8 Gy of irradiation, mitotic chromosome spreads were prepared, and chromosome aberrations were scored from 40 independent metaphase cells as described in Materials and Methods. Data shown are averages ± SE (Standard Error). The Chi-Square Test was used to calculate the p-values, which are shown in the figure. Panel D shows the rate of micronuclear formations in A7 and M2 cells. Cytokinesis was blocked by treatment with cytochalasin B to form binuclear cells 24 h after γ-irradiation. Micronuclei in binuclear cells were visualized by Giemsa staining and scored under light microscopy. About 1000 binuclear cells in each experiment were analyzed and the data are averages of 5–6 independent experiments. Shown is the percentage of binuclear cells that have at least one micronucleus. The p-values indicate statistic significance based on t -tests.
Figure Legend Snippet: Filamin-A deficiency impairs the DNA double strand break repair. Panel A shows the numbers of γH2AX foci per cell in A7 and M2 cells. At time points after the cells were treated with γ-radiation (panel A), cells were fixed and stained by immunoflurescent techniques with an anti-γH2AX antibody. The number of γH2AX foci was counted in > 400 cells per time point in each experiment. Data shown are averages and standard errors of three experiments. Panel B shows the total γH2AX protein level measured by anti-γH2AX western blot at time points after irradiation (4 Gy) in C8161 and MB231 cells. The dose and times after irradiation are shown on the top of the panel. “FLNa-shRNA” indicates the whether the cells express filamin-A-specific shRNA. Beta-actin was used as a loading control. Panel C shows the total number of unstable chromosome aberrations per metaphase cell in A7 and M2. At 15 min, 8 hours, and 24 hours after 8 Gy of irradiation, mitotic chromosome spreads were prepared, and chromosome aberrations were scored from 40 independent metaphase cells as described in Materials and Methods. Data shown are averages ± SE (Standard Error). The Chi-Square Test was used to calculate the p-values, which are shown in the figure. Panel D shows the rate of micronuclear formations in A7 and M2 cells. Cytokinesis was blocked by treatment with cytochalasin B to form binuclear cells 24 h after γ-irradiation. Micronuclei in binuclear cells were visualized by Giemsa staining and scored under light microscopy. About 1000 binuclear cells in each experiment were analyzed and the data are averages of 5–6 independent experiments. Shown is the percentage of binuclear cells that have at least one micronucleus. The p-values indicate statistic significance based on t -tests.

Techniques Used: Staining, Western Blot, Irradiation, shRNA, Light Microscopy

Knockdown of filamin-A inhibits homologous recombinational repair of DSB. The filamin-A expression in HT1080–1885 cells, which host an HR assay substrate, was knocked down by shRNA. Five control clones (labeled as 1–5) and seven knockdown clones (labeled as A-G) were isolated. Break-induced HR was measured in these clones (See Results for a description of the HR assay). Panel A shows the level of filamin-A protein levels in control and knockdown clones. Clones 1–5 are cells expressing a control shRNA, and clones A-G are cells expressing filamin-A specific shRNA. HA-ISceI blot (bottom panel) shows the expression level of the I-SceI enzymes that introduces the site specific DSB in the HT1080–1885 cells. Panel B illustrates the recombination substrate and the HR mechanism resulting in the reconstitution of a functional puromycin resistance gene. Panel C shows the HR frequencies of individual clones. Shown are means and standard deviations of 6–8 experiments for each clone. Panel D summarizes HR frequencies for the filamin-A knockdown clones and the control clones. Shown are the means and standard deviations. The p-value indicates the statistical significance of the difference between means of the control clones and the filamin-A knock down clones ( t -test).
Figure Legend Snippet: Knockdown of filamin-A inhibits homologous recombinational repair of DSB. The filamin-A expression in HT1080–1885 cells, which host an HR assay substrate, was knocked down by shRNA. Five control clones (labeled as 1–5) and seven knockdown clones (labeled as A-G) were isolated. Break-induced HR was measured in these clones (See Results for a description of the HR assay). Panel A shows the level of filamin-A protein levels in control and knockdown clones. Clones 1–5 are cells expressing a control shRNA, and clones A-G are cells expressing filamin-A specific shRNA. HA-ISceI blot (bottom panel) shows the expression level of the I-SceI enzymes that introduces the site specific DSB in the HT1080–1885 cells. Panel B illustrates the recombination substrate and the HR mechanism resulting in the reconstitution of a functional puromycin resistance gene. Panel C shows the HR frequencies of individual clones. Shown are means and standard deviations of 6–8 experiments for each clone. Panel D summarizes HR frequencies for the filamin-A knockdown clones and the control clones. Shown are the means and standard deviations. The p-value indicates the statistical significance of the difference between means of the control clones and the filamin-A knock down clones ( t -test).

Techniques Used: Expressing, shRNA, Clone Assay, Labeling, Isolation, Functional Assay

Filamin-A truncations fail to complement the DSB repair defects. Several truncation mutants of filamin-A as illustrated in 6A (bottom panel) was expressed in the M2 filamin-A deficient cells. See text for full description of the truncation mutants. The cell line expressing the full length filamin-A was used as positive control, and cell line carrying an empty vector was used as negative control. Panel B shows the average number of γH2AX foci per cells shortly after irradiation (0.25 hr), and after 2 and 8 hours of repair for M2-Vector (negative control), A7 (M2 cells with full length filamin-A), and several M2 cells expressing truncated filamin-A . Panel C are Western blots used as an alterative method to measure the level of γH2AX at various times after irradiation among the same cell lines. Anti-Actin blot was used as loading control for γH2AX blot. Panel C (bottom panel) also shows the confirmed expression of filamin-A truncation mutants.
Figure Legend Snippet: Filamin-A truncations fail to complement the DSB repair defects. Several truncation mutants of filamin-A as illustrated in 6A (bottom panel) was expressed in the M2 filamin-A deficient cells. See text for full description of the truncation mutants. The cell line expressing the full length filamin-A was used as positive control, and cell line carrying an empty vector was used as negative control. Panel B shows the average number of γH2AX foci per cells shortly after irradiation (0.25 hr), and after 2 and 8 hours of repair for M2-Vector (negative control), A7 (M2 cells with full length filamin-A), and several M2 cells expressing truncated filamin-A . Panel C are Western blots used as an alterative method to measure the level of γH2AX at various times after irradiation among the same cell lines. Anti-Actin blot was used as loading control for γH2AX blot. Panel C (bottom panel) also shows the confirmed expression of filamin-A truncation mutants.

Techniques Used: Expressing, Positive Control, Plasmid Preparation, Negative Control, Irradiation, Western Blot

); 2) C8161 (control shRNA) and C8161-KD (FLNa-shRNA): C8161 is filamin-A positive metastatic melanoma cell line expressing a control shRNA, and C8161-KD is a C8161 sub-clone expressing a filamin-A specific shRNA; 3) MB231 (control shRNA) and MB231-KD (FLNa-shRNA): MB231 is the MDA-MB-231 breast cancer cell line expressing a control shRNA, and MB231-KD is a MDA-MB-231 sub-clone expressing a filamin-A specific shRNA. Beta-actin was used as a loading control. Colony formation assays (see Materials and Methods) were used to determine the survival fractions after irradiation. Panel B-C are the radiation survival curves for the three pairs of filamin-A proficient and deficient cells. Error bars represent the standard deviation of three independent experiments.
Figure Legend Snippet: ); 2) C8161 (control shRNA) and C8161-KD (FLNa-shRNA): C8161 is filamin-A positive metastatic melanoma cell line expressing a control shRNA, and C8161-KD is a C8161 sub-clone expressing a filamin-A specific shRNA; 3) MB231 (control shRNA) and MB231-KD (FLNa-shRNA): MB231 is the MDA-MB-231 breast cancer cell line expressing a control shRNA, and MB231-KD is a MDA-MB-231 sub-clone expressing a filamin-A specific shRNA. Beta-actin was used as a loading control. Colony formation assays (see Materials and Methods) were used to determine the survival fractions after irradiation. Panel B-C are the radiation survival curves for the three pairs of filamin-A proficient and deficient cells. Error bars represent the standard deviation of three independent experiments.

Techniques Used: shRNA, Expressing, Multiple Displacement Amplification, Irradiation, Standard Deviation

Related Articles

Western Blot:

Article Title: Filamin-A as a marker and target for DNA damage based cancer therapy
Article Snippet: .. The antibodies for filamin-A (Chemicon International, Inc., CA), RAD51 (Santa Cruz Biotechnologh, Inc., Santa Cruz, CA), and γ H2AX-Ser139 (Upstate Biotechnology, Lake Placid, NY) were used for the Western blot. .. The β-actin level was detected with anti-actin antibody (Sigma-Aldrich, St. Louis, MO) to serve as a loading control for whole protein extract.

Article Title: EHD2 is a mechanotransducer connecting caveolae dynamics with gene transcription
Article Snippet: .. Antibodies and reagents The following commercially available antibodies were used for Western blotting: mouse monoclonal antibodies against clathrin heavy chain (CHC; BD Biosciences; 610500), lamin A/C (Santa Cruz Biotechnology; sc-7292), Hsp90 (Santa Cruz Biotechnology; sc-13119), EHD2 (Santa Cruz Biotechnology; sc-100724), dynamin (BD Biosciences; 610245), and filamin A (Chemicon; MAB1678); rabbit polyclonal antibodies against SUMO2/3 (Cell Signaling Technology; 4971), Cav1 (BD Biosciences; 610059), pacsin2 (Abgent; AP8088b); and cavin1 (Sigma; AV36965); for immunofluorescence, mouse monoclonal anti-cavin1 (BD Biosciences; 611258), goat polyclonal anti-EHD2 (Abcam; Ab23935), rabbit polyclonal anti-Cav1 (BD Biosciences; 610059), mouse monoclonal anti-lamin A/C (Santa Cruz Biotechnology; sc-7292), rabbit polyclonal anti-SUMO1 (Cell Signaling Technology; 4930), and rabbit polyclonal anti-SUMO2/3 (Cell Signaling Technology; 4971). .. Antibodies conjugated to Alexa Fluor 488, Cy3, Cy5, or HRP (Beckman Coulter and Invitrogen) were used as secondary antibodies.

Incubation:

Article Title: Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility
Article Snippet: .. Biotinylated Wnt5A (5 μg/ml, R & D), Klotho (5 μg/ml, Abcam, Cambridge, MA), Filamin A (1:100, Millipore, Billerica, MA), syndecan-1 (1:100, R & D), and syndecan-4 (1:100, R & D) primary antibodies were diluted in blocking buffer, and slides were incubated overnight at 4°C. .. Cells were washed in PBS and incubated with Alexa Fluor-568 (1:2,000) or Alexa-488 (1:2,000) conjugated secondary antibodies (Invitrogen) in blocking buffer for 1 hour at room temperature.

Immunofluorescence:

Article Title: EHD2 is a mechanotransducer connecting caveolae dynamics with gene transcription
Article Snippet: .. Antibodies and reagents The following commercially available antibodies were used for Western blotting: mouse monoclonal antibodies against clathrin heavy chain (CHC; BD Biosciences; 610500), lamin A/C (Santa Cruz Biotechnology; sc-7292), Hsp90 (Santa Cruz Biotechnology; sc-13119), EHD2 (Santa Cruz Biotechnology; sc-100724), dynamin (BD Biosciences; 610245), and filamin A (Chemicon; MAB1678); rabbit polyclonal antibodies against SUMO2/3 (Cell Signaling Technology; 4971), Cav1 (BD Biosciences; 610059), pacsin2 (Abgent; AP8088b); and cavin1 (Sigma; AV36965); for immunofluorescence, mouse monoclonal anti-cavin1 (BD Biosciences; 611258), goat polyclonal anti-EHD2 (Abcam; Ab23935), rabbit polyclonal anti-Cav1 (BD Biosciences; 610059), mouse monoclonal anti-lamin A/C (Santa Cruz Biotechnology; sc-7292), rabbit polyclonal anti-SUMO1 (Cell Signaling Technology; 4930), and rabbit polyclonal anti-SUMO2/3 (Cell Signaling Technology; 4971). .. Antibodies conjugated to Alexa Fluor 488, Cy3, Cy5, or HRP (Beckman Coulter and Invitrogen) were used as secondary antibodies.

Blocking Assay:

Article Title: Loss Of Klotho During Melanoma Progression Leads To Increased Filamin Cleavage, Increased Wnt5A Expression and Enhanced Melanoma Cell Motility
Article Snippet: .. Biotinylated Wnt5A (5 μg/ml, R & D), Klotho (5 μg/ml, Abcam, Cambridge, MA), Filamin A (1:100, Millipore, Billerica, MA), syndecan-1 (1:100, R & D), and syndecan-4 (1:100, R & D) primary antibodies were diluted in blocking buffer, and slides were incubated overnight at 4°C. .. Cells were washed in PBS and incubated with Alexa Fluor-568 (1:2,000) or Alexa-488 (1:2,000) conjugated secondary antibodies (Invitrogen) in blocking buffer for 1 hour at room temperature.

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    Millipore filamin a
    Disrupting the <t>CaR-filamin</t> interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin
    Filamin A, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin

    Journal: The Journal of investigative dermatology

    Article Title: The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A

    doi: 10.1038/jid.2010.414

    Figure Lengend Snippet: Disrupting the CaR-filamin interaction blocked the Ca 2+ o -induced Rho activation and membrane complex formation of CaR, Rho A and E-cadherin

    Article Snippet: The following antibodies (Abs) were used in this study: monoclonal antibodies (mAbs) for α2 integrin (BD Biosciences, San Jose, CA), for human involucrin and β-actin, for c-Myc- and HA-tags (Sigma-Aldrich, St. Louis, MO), polyclonal and monoclonal Abs against E-cadherin, β-, γ-, and p120-catenin and Fyn, polyclonal Abs against Rho A, Rac1 and cdc42, and CaR, mAb against transglutaminase 1 (Santa Cruz Biotechnology, Santa Cruz, CA), mAbs for phosphotyrosine, Rho and filamin A (Millipore Corp., Temecula, CA), polyclonal Abs for human keratin 1, keratin 14 and loricrin (Covance Inc., Berkeley, CA), and rabbit anti-human CaR ( ).

    Techniques: Activation Assay

    Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane

    Journal: The Journal of investigative dermatology

    Article Title: The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A

    doi: 10.1038/jid.2010.414

    Figure Lengend Snippet: Elevated [Ca 2+ ] o induced complex formation of CaR, , Rho A, filamin A and E-cadherin at the cell membrane

    Article Snippet: The following antibodies (Abs) were used in this study: monoclonal antibodies (mAbs) for α2 integrin (BD Biosciences, San Jose, CA), for human involucrin and β-actin, for c-Myc- and HA-tags (Sigma-Aldrich, St. Louis, MO), polyclonal and monoclonal Abs against E-cadherin, β-, γ-, and p120-catenin and Fyn, polyclonal Abs against Rho A, Rac1 and cdc42, and CaR, mAb against transglutaminase 1 (Santa Cruz Biotechnology, Santa Cruz, CA), mAbs for phosphotyrosine, Rho and filamin A (Millipore Corp., Temecula, CA), polyclonal Abs for human keratin 1, keratin 14 and loricrin (Covance Inc., Berkeley, CA), and rabbit anti-human CaR ( ).

    Techniques:

    Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions

    Journal: The Journal of investigative dermatology

    Article Title: The calcium-sensing receptor-dependent regulation of cell-cell adhesion and keratinocyte differentiation requires Rho and Filamin A

    doi: 10.1038/jid.2010.414

    Figure Lengend Snippet: Interfering with the CaR-filamin interaction suppressed Ca 2+ o -induced formation of adherens junctions

    Article Snippet: The following antibodies (Abs) were used in this study: monoclonal antibodies (mAbs) for α2 integrin (BD Biosciences, San Jose, CA), for human involucrin and β-actin, for c-Myc- and HA-tags (Sigma-Aldrich, St. Louis, MO), polyclonal and monoclonal Abs against E-cadherin, β-, γ-, and p120-catenin and Fyn, polyclonal Abs against Rho A, Rac1 and cdc42, and CaR, mAb against transglutaminase 1 (Santa Cruz Biotechnology, Santa Cruz, CA), mAbs for phosphotyrosine, Rho and filamin A (Millipore Corp., Temecula, CA), polyclonal Abs for human keratin 1, keratin 14 and loricrin (Covance Inc., Berkeley, CA), and rabbit anti-human CaR ( ).

    Techniques:

    Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

    Journal: Journal of dermatological science

    Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

    doi: 10.1016/j.jdermsci.2013.09.007

    Figure Lengend Snippet: Inhibition of filamin A expression by siRNA blocked the Ca 2+ o -induced formation of a signaling protein complex at the cell membrane and activation of Rho. Keratinocytes were transfected with filamin A-specific (siFLNA), Trio-specific (siTrio) or control

    Article Snippet: Analyses of immunoprecipitates showed that Ca2+ o increased the protein level of Rho A in the cell membrane as well as the amount of associated CaR, filamin A, Trio, and E-cadherin in control keratinocytes ( ).

    Techniques: Inhibition, Expressing, Activation Assay, Transfection

    Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

    Journal: Journal of dermatological science

    Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

    doi: 10.1016/j.jdermsci.2013.09.007

    Figure Lengend Snippet: Expressing dominate-negative peptides that disrupt filamin-Trio and filamin-Rho interactions inhibited Ca 2+ o -induced activation of Rho A and Rac1. Keratinocytes were infected by adenoviruses expressing the Trio GEFD1 PH (TrioPH1), Trio GEFD2 PH (TrioPH2)

    Article Snippet: Analyses of immunoprecipitates showed that Ca2+ o increased the protein level of Rho A in the cell membrane as well as the amount of associated CaR, filamin A, Trio, and E-cadherin in control keratinocytes ( ).

    Techniques: Expressing, Activation Assay, Infection

    Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

    Journal: Journal of dermatological science

    Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

    doi: 10.1016/j.jdermsci.2013.09.007

    Figure Lengend Snippet: Inhibition of filamin A or Trio expression suppressed Ca 2+ o -induced colocalization of β-catenin, Rho A and E-cadherin at the cell-cell junctions. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes

    Article Snippet: Analyses of immunoprecipitates showed that Ca2+ o increased the protein level of Rho A in the cell membrane as well as the amount of associated CaR, filamin A, Trio, and E-cadherin in control keratinocytes ( ).

    Techniques: Inhibition, Expressing, Transfection

    Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

    Journal: Journal of dermatological science

    Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

    doi: 10.1016/j.jdermsci.2013.09.007

    Figure Lengend Snippet: Elevated [Ca 2+ ] o induced colocalization of Rho A, CaR, filamin A and E-cadherin at the cell-cell junctions. Keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes to induce cell-cell contacts. Fluorescence immunostaining was performed using antibodies

    Article Snippet: Analyses of immunoprecipitates showed that Ca2+ o increased the protein level of Rho A in the cell membrane as well as the amount of associated CaR, filamin A, Trio, and E-cadherin in control keratinocytes ( ).

    Techniques: Fluorescence, Immunostaining

    Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

    Journal: Journal of dermatological science

    Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

    doi: 10.1016/j.jdermsci.2013.09.007

    Figure Lengend Snippet: Filamin A serves as a scaffold for the assembly of a signaling complex required for E-cadherin mediated cell-cell adhesion. In response to Ca stimulation, CaR, Trio, and Rho A form a signaling complex at the cell membrane to activate downstream effectors

    Article Snippet: Analyses of immunoprecipitates showed that Ca2+ o increased the protein level of Rho A in the cell membrane as well as the amount of associated CaR, filamin A, Trio, and E-cadherin in control keratinocytes ( ).

    Techniques:

    Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

    Journal: Journal of dermatological science

    Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

    doi: 10.1016/j.jdermsci.2013.09.007

    Figure Lengend Snippet: Inhibition of filamin A or Trio expression suppressed keratinocyte cell-cell adhesion. SiFLNA-, siTrio- or siControl-transfected keratinocytes were exposed to 2 mM Ca 2+ for 10 minutes. Plasma membrane lysates were immunoprecipitated (IP) with E-cadherin

    Article Snippet: Analyses of immunoprecipitates showed that Ca2+ o increased the protein level of Rho A in the cell membrane as well as the amount of associated CaR, filamin A, Trio, and E-cadherin in control keratinocytes ( ).

    Techniques: Inhibition, Expressing, Transfection, Immunoprecipitation

    Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

    Journal: Journal of dermatological science

    Article Title: Obligatory roles of filamin A in E-cadherin-mediated cell-cell adhesion in epidermal keratinocytes

    doi: 10.1016/j.jdermsci.2013.09.007

    Figure Lengend Snippet: Disrupting filamin-Trio and filamin-Rho interactions decreased the Ca 2+ o -induced formation of a protein complex containing filamin, Trio, Rho A and E-cadherin at the cell membrane and suppressed keratinocyte cell-cell adhesion. Keratinocytes were infected

    Article Snippet: Analyses of immunoprecipitates showed that Ca2+ o increased the protein level of Rho A in the cell membrane as well as the amount of associated CaR, filamin A, Trio, and E-cadherin in control keratinocytes ( ).

    Techniques: Infection