Fig. 2

CRN5 localises to the nucleus, F-actin stress fibres, and lamellipodia. a Cytosolic and nuclear (arrows) localisation of CRN5 in CT26 mouse colon carcinoma cells. Detection was done with pAb H-110. Cells were fixed with 4 % paraformaldehyde, permeabilised with Triton X-100, and treated with 0.05 % Trypsin for 10 min for epitope retrieval according to [25]. Secondary antibody, goat anti-mouse AlexaFluor 488; nuclei were visualized with DAPI. b Nuclei of HeLa cells over-expressing FLAG-tagged CRN5 were isolated, and presence of CRN5 was determined by pAb H-110 immunoblotting. For control, the same membrane was labelled with lamin B1 (nuclear envelope) and α-tubulin (cytoplasm) antibodies. c In A7r5 aortic smooth muscle cells GFP-CRN5 localised to F-actin stress fibres; arrows label a cell-cell contact. F-actin was visualised using TRITC-phalloidin. d In U373 human glioblastoma cells GFP-CRN5 co-localised with the Arp2/3 complex (anti-p34 antibody) at the front of lamellipodia (arrows). e, f U373 human glioblastoma cells over-expressing GFP-CRN5 or GFP-CRN2. While GFP-CRN2 was strongly enriched at all F-actin structures including stress fibres, the presence of GFP-CRN5 led to depletion of F-actin stress fibres (asterisk) and induced extensions at the rear of the cells (arrowheads)