f actin reporter  (Addgene inc)

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cancer cells suppress NK cell activity by actin-driven polarization of inhibitory ligands to the immunological synapse

doi: 10.1073/pnas.2503259122

Figure Lengend Snippet: Live-cell confocal time-lapse of Emerald-LifeAct-expressing MDA-MB-231 cells (green) co-cultured with CMRA-labeled primary NK (pNK) cells (red) in medium containing SYTOX Blue dye (cyan).The right panel shows F-actin rendered in pseudocolor. The sequence captures polarized F-actin remodelling at the immunological synapse during repeated contacts with pNK cells on one side of the target cell. The imaged cells are the same as those depicted in Fig. S1A.

Article Snippet: Breast cancer cell lines were transduced to stably express the mEmerald-Lifeact-7 F-actin reporter (Addgene, plasmid #54148), as previously described ( ).

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cancer cells suppress NK cell activity by actin-driven polarization of inhibitory ligands to the immunological synapse

doi: 10.1073/pnas.2503259122

Figure Lengend Snippet: Live-cell confocal time-lapse of Emerald-LifeAct-expressing MDA-MB-231 cells (green) co-cultured with CMRA-labelled primary NK (pNK) cells (red) in medium containing SYTOX Blue (cyan). The right panel shows F-actin rendered in pseudocolour. The sequence shows two target cells: one engages in a brief pNK-cell contact, while the other maintains a sustained ≈1 h contact with continuous F-actin remodelling. The imaged cells are the same as those depicted in Fig. S1B.

Article Snippet: Breast cancer cell lines were transduced to stably express the mEmerald-Lifeact-7 F-actin reporter (Addgene, plasmid #54148), as previously described ( ).

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cancer cells suppress NK cell activity by actin-driven polarization of inhibitory ligands to the immunological synapse

doi: 10.1073/pnas.2503259122

Figure Lengend Snippet: Live-cell confocal time-lapse of Emerald-LifeAct-expressing MDA-MB-231 cells (green) co-cultured with CMRA-labelled primary NK (pNK) cells (red) in medium containing SYTOX Blue (cyan). The right panel shows F-actin rendered in pseudocolour. The sequence captures a target cell displaying F-actin remodelling during sequential contacts with three pNK cells. The imaged cells are the same as those depicted in Fig. S1C.

Article Snippet: Breast cancer cell lines were transduced to stably express the mEmerald-Lifeact-7 F-actin reporter (Addgene, plasmid #54148), as previously described ( ).

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cancer cells suppress NK cell activity by actin-driven polarization of inhibitory ligands to the immunological synapse

doi: 10.1073/pnas.2503259122

Figure Lengend Snippet: Live-cell confocal time-lapse of Emerald-LifeAct-expressing MDA-MB-231 cells (green) co-cultured with CMRA-labelled primary NK (pNK) cells (red) in medium containing SYTOX Blue (cyan). The right panel shows F-actin rendered in pseudocolour. The sequence shows a target cell that lacks F-actin remodelling at the immunological synapse and undergoes rapid cell death after pNK-cell contact. The imaged cells are the same as those depicted in Fig. S1D.

Article Snippet: Breast cancer cell lines were transduced to stably express the mEmerald-Lifeact-7 F-actin reporter (Addgene, plasmid #54148), as previously described ( ).

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cancer cells suppress NK cell activity by actin-driven polarization of inhibitory ligands to the immunological synapse

doi: 10.1073/pnas.2503259122

Figure Lengend Snippet: Live-cell confocal time-lapse of Emerald-LifeAct-expressing MDA-MB-231 cells (green) co-cultured with CMRA-labelled primary NK (pNK) cells (red) in medium containing SYTOX Blue (cyan). The right panel shows F-actin rendered in pseudocolour. The sequence shows a target cell that lacks F-actin remodelling at the immunological synapse and undergoes rapid cell death after pNK-cell contact. The imaged cells are the same as those depicted in Fig. S1E.

Article Snippet: Breast cancer cell lines were transduced to stably express the mEmerald-Lifeact-7 F-actin reporter (Addgene, plasmid #54148), as previously described ( ).

Techniques:

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Cancer cells suppress NK cell activity by actin-driven polarization of inhibitory ligands to the immunological synapse

doi: 10.1073/pnas.2503259122

Figure Lengend Snippet: Live-cell confocal time-lapse of Emerald-LifeAct-expressing MDA-MB-231 cells (green) co-cultured with CMRA-labelled primary NK (pNK) cells (red) in medium containing SYTOX Blue (cyan). The right panel shows F-actin rendered in pseudocolour. The sequence shows a target cell that lacks F-actin remodelling at the immunological synapse and undergoes rapid cell death after pNK-cell contact. The imaged cells are the same as those depicted in Fig. S1F.

Article Snippet: Breast cancer cell lines were transduced to stably express the mEmerald-Lifeact-7 F-actin reporter (Addgene, plasmid #54148), as previously described ( ).

Techniques:

Journal: eLife

Article Title: Epithelial cell chirality emerges through the dynamic concentric pattern of actomyosin cytoskeleton

doi: 10.7554/eLife.102296

Figure Lengend Snippet: Fluorescent images of Caco-2 expressing Lifeact-mEmelard taken by lattice light-sheet microscopy (LLSM) at three different z -planes ( z = 0, 0.5, and 1.0 µm).

Article Snippet: The construction of pLVSIN-EF1a-IRES-pur has been described previously ( ). mEmerald-Lifeact-7 was a gift from Michael Davidson (Addgene plasmid # 54148; http://n2t.net/addgene :54148; RRID: Addgene_54148 ).

Techniques: