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. 2015 Mar 2;5(7):656–666. doi: 10.7150/thno.10844

Figure 4.

Figure 4

CYLD enhances noscapine activity to induce mitotic arrest and apoptosis in a microtubule-dependent manner. (A) Molt4 cells were transfected with GFP, GFP-CYLD, GFP-CYLD-ΔCG1, or GFP-CYLD-C/S and treated with 2 μM noscapine for 36 hours, followed by staining with PI (red). (B) Experiments were performed as in A, and the percentage of apoptotic cells (with apoptotic bodies) was quantified. (C) Molt4 cells were transfected with GFP, GFP-CYLD, GFP-CYLD-ΔCG1, or GFP-CYLD-C/S and treated with 2 μM noscapine for 24 hours. Cells were then stained with PI (red). (D) Experiments were performed as in C, and the percentage of mitotic cells (with chromosome rosettes) was quantified. (E) Molt4 cells were transfected with GFP, GFP-CYLD, GFP-CYLD-ΔCG1, or GFP-CYLD-C/S and treated with 2 μM noscapine for 36 hours. Cells were then stained with PI and Alexa Fluor 488-conjugated annexin V, and analyzed by flow cytometry. (F) Experiments were performed as in E, and the percentage of annexin V-positive cells was quantified. (G) Molt4 cells were transfected with GFP, GFP-CYLD, GFP-CYLD-ΔCG1, or GFP-CYLD-C/S and treated with 2 μM noscapine for 24 hours. Cells were then stained with PI and analyzed by flow cytometry. (H) Experiments were performed as in G, and the percentage of cells in sub-G1, G1/S, and G2/M phases were determined. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, not significant.