Figure 3
From: Foci of cyclin A2 interact with actin and RhoA in mitosis
Cyclin A2 interacts with actin in mitosis.
(A) U2OS cells were synchronised in G1/S or M, treated with latrunculin B or DMSO and fractionated. Immunoblots showing cyclin A2 and actin levels in the soluble (SN: supernatant) and insoluble (P: pellet) fractions. GAPDH is used as a loading control. (B) Spin-down assay with F-actin using purified cyclin A2-GST. α-actinin is used as positive control, BSA and GST as negative controls. Coomassie blue staining following SDS-PAGE. Data shown in (A,B) are representative of three independent experiments. (C) Left panels, live MCF-7 cells microinjected with pEGFP-N1-cyclin A2, with or without pmCherry-C1-β-actin. The co-injected cell is followed through time. Two-photon EGFP images and corresponding EGFP lifetime maps. mCherry confocal images. Representative images of 10 cells (100% showing the same transient interaction) analysed in two independent experiments. Scale bars: 2 μm. Right panel, data represent the mean ± s.e.m. of EGFP lifetimes in cyclin A2-EGFP foci (n = 40, P < 0.0005).
