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(A) Schematic of Mitotrap used in this experiment. In all cases cells expressing endogenously tagged GFP-CLIC4 were co-transfected with Cry2-GFP-VHH and <t>CIB-Tom20</t> plasmids and pulsed with a 488nm laser to re-target GFP-CLIC4 at the mitochondria. (B) Interphase cell exposed to 488nm to activate Mitotrap. Mitochondria is labeled in red and endogenous GFP-CLIC4 in green. (C-D) Still images from time-lapse microscopy where Mitotrap was activated. Arrows point to blebs or cytokinesis failure induced by GFP-CLIC4 Mitotrap. (E) Quantification of time required for cells to complete mitotic cell division in control and Mitotrapped cells. Data shown are the means and standard deviations.
Tom20 Cib Stop, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A) Schematic of Mitotrap used in this experiment. In all cases cells expressing endogenously tagged GFP-CLIC4 were co-transfected with Cry2-GFP-VHH and CIB-Tom20 plasmids and pulsed with a 488nm laser to re-target GFP-CLIC4 at the mitochondria. (B) Interphase cell exposed to 488nm to activate Mitotrap. Mitochondria is labeled in red and endogenous GFP-CLIC4 in green. (C-D) Still images from time-lapse microscopy where Mitotrap was activated. Arrows point to blebs or cytokinesis failure induced by GFP-CLIC4 Mitotrap. (E) Quantification of time required for cells to complete mitotic cell division in control and Mitotrapped cells. Data shown are the means and standard deviations.

Journal: bioRxiv

Article Title: CLIC4 is a cytokinetic cleavage furrow protein that regulates cortical cytoskeleton stability during cell division

doi: 10.1101/723940

Figure Lengend Snippet: (A) Schematic of Mitotrap used in this experiment. In all cases cells expressing endogenously tagged GFP-CLIC4 were co-transfected with Cry2-GFP-VHH and CIB-Tom20 plasmids and pulsed with a 488nm laser to re-target GFP-CLIC4 at the mitochondria. (B) Interphase cell exposed to 488nm to activate Mitotrap. Mitochondria is labeled in red and endogenous GFP-CLIC4 in green. (C-D) Still images from time-lapse microscopy where Mitotrap was activated. Arrows point to blebs or cytokinesis failure induced by GFP-CLIC4 Mitotrap. (E) Quantification of time required for cells to complete mitotic cell division in control and Mitotrapped cells. Data shown are the means and standard deviations.

Article Snippet: For the Mitotrap experiments, cells expressing endogenous GFP-CLIC4 were transfected with Cry2-VHH Addgene #58370) and Tom20-CIB-stop (Addgene #117243).

Techniques: Expressing, Transfection, Labeling, Time-lapse Microscopy