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argent regulated homodimerization kit pc 4 -fv 1 e  (ARIAD Inc)
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ARIAD Inc argent regulated homodimerization kit pc 4 -fv 1 e
( A ) Illustration of <t>FKBP-dimer</t> system. Protein of interest is tagged with FKBP domain. Cell-permeable agent B/B homodimerizer (dimerizer) forces FKBP domains together. ( B ) Cultured keratinocytes were transfected with Celsr1 Crsh -FKBP-HA, switched to 1.5 mM calcium for 24 hr, and treated with ethanol (control) or dimerizer for 2 hr prior to the end of the calcium switch to induce dimerization of the FKBP domain fused to Celsr1 Crsh . Yellow arrows highlight position of the cell–cell interface between Celsr1 Crsh -FKBP-HA expressing cells. ( C ) Border enrichment quantification for Celsr1 Crsh -FKBP-HA. Ethanol, n = 14; Dimerizer, n = 12. One-tailed, unpaired t -test, **p=0.0057. ( D ) Cell mixing assay where cells were transfected separately with either Celsr1 WT -GFP or Celsr1 Crsh -FKBP-HA and mixed. Shown are representative examples of mixed-cell junctions between Celsr1 WT -GFP (top cell in pair, green) and Celsr1 Crsh -FKBP-HA (bottom cell in pair, red) treated with ethanol (control) or dimerizer. Scale bars, 10 µm. Figure 6—source data 1. Data accompanying .
Argent Regulated Homodimerization Kit Pc 4 Fv 1 E, supplied by ARIAD Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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( A ) Illustration of FKBP-dimer system. Protein of interest is tagged with FKBP domain. Cell-permeable agent B/B homodimerizer (dimerizer) forces FKBP domains together. ( B ) Cultured keratinocytes were transfected with Celsr1 Crsh -FKBP-HA, switched to 1.5 mM calcium for 24 hr, and treated with ethanol (control) or dimerizer for 2 hr prior to the end of the calcium switch to induce dimerization of the FKBP domain fused to Celsr1 Crsh . Yellow arrows highlight position of the cell–cell interface between Celsr1 Crsh -FKBP-HA expressing cells. ( C ) Border enrichment quantification for Celsr1 Crsh -FKBP-HA. Ethanol, n = 14; Dimerizer, n = 12. One-tailed, unpaired t -test, **p=0.0057. ( D ) Cell mixing assay where cells were transfected separately with either Celsr1 WT -GFP or Celsr1 Crsh -FKBP-HA and mixed. Shown are representative examples of mixed-cell junctions between Celsr1 WT -GFP (top cell in pair, green) and Celsr1 Crsh -FKBP-HA (bottom cell in pair, red) treated with ethanol (control) or dimerizer. Scale bars, 10 µm. Figure 6—source data 1. Data accompanying .

Journal: eLife

Article Title: Celsr1 adhesive interactions mediate the asymmetric organization of planar polarity complexes

doi: 10.7554/eLife.62097

Figure Lengend Snippet: ( A ) Illustration of FKBP-dimer system. Protein of interest is tagged with FKBP domain. Cell-permeable agent B/B homodimerizer (dimerizer) forces FKBP domains together. ( B ) Cultured keratinocytes were transfected with Celsr1 Crsh -FKBP-HA, switched to 1.5 mM calcium for 24 hr, and treated with ethanol (control) or dimerizer for 2 hr prior to the end of the calcium switch to induce dimerization of the FKBP domain fused to Celsr1 Crsh . Yellow arrows highlight position of the cell–cell interface between Celsr1 Crsh -FKBP-HA expressing cells. ( C ) Border enrichment quantification for Celsr1 Crsh -FKBP-HA. Ethanol, n = 14; Dimerizer, n = 12. One-tailed, unpaired t -test, **p=0.0057. ( D ) Cell mixing assay where cells were transfected separately with either Celsr1 WT -GFP or Celsr1 Crsh -FKBP-HA and mixed. Shown are representative examples of mixed-cell junctions between Celsr1 WT -GFP (top cell in pair, green) and Celsr1 Crsh -FKBP-HA (bottom cell in pair, red) treated with ethanol (control) or dimerizer. Scale bars, 10 µm. Figure 6—source data 1. Data accompanying .

Article Snippet: Celsr1 Crsh -FKBP-HA was generated by replacing EGFP in Celsr1 Crsh -GFP ( ) with HA-tagged FKBP (pC 4 -Fv 1 E; ARGENT Regulated Homodimerization Kit, Ariad) digested with HindIII and NotI sites.

Techniques: Cell Culture, Transfection, Control, Expressing, One-tailed Test

Additional representative examples of cell borders of neighboring cells both expressing transfected constructs from cis -dimerization FBKP experiment. Images cropped and rotated to display border with horizontal orientation. Mouse keratinocytes were transfected with constructs indicated, switched to 1.5 mM calcium for 24 hr along with treatment of either ethanol (control) or dimerizer (B/B homo-dimerizer) for the final 2 hr of the calcium switch to induce FKBP domain dimerization. ( A ) Transfection of just Celsr1 Crsh -FKBP-HA. ( B ) Co-transfection of both Celsr1 WT -GFP with Celsr1 Crsh -FKBP-HA. ( C ) Mixed culture of cells transfected with either Celsr1 WT -GFP or Celsr1 Crsh -FKBP-HA. Scale bar, 10 µm.

Journal: eLife

Article Title: Celsr1 adhesive interactions mediate the asymmetric organization of planar polarity complexes

doi: 10.7554/eLife.62097

Figure Lengend Snippet: Additional representative examples of cell borders of neighboring cells both expressing transfected constructs from cis -dimerization FBKP experiment. Images cropped and rotated to display border with horizontal orientation. Mouse keratinocytes were transfected with constructs indicated, switched to 1.5 mM calcium for 24 hr along with treatment of either ethanol (control) or dimerizer (B/B homo-dimerizer) for the final 2 hr of the calcium switch to induce FKBP domain dimerization. ( A ) Transfection of just Celsr1 Crsh -FKBP-HA. ( B ) Co-transfection of both Celsr1 WT -GFP with Celsr1 Crsh -FKBP-HA. ( C ) Mixed culture of cells transfected with either Celsr1 WT -GFP or Celsr1 Crsh -FKBP-HA. Scale bar, 10 µm.

Article Snippet: Celsr1 Crsh -FKBP-HA was generated by replacing EGFP in Celsr1 Crsh -GFP ( ) with HA-tagged FKBP (pC 4 -Fv 1 E; ARGENT Regulated Homodimerization Kit, Ariad) digested with HindIII and NotI sites.

Techniques: Expressing, Transfection, Construct, Control, Cotransfection