Figure 5

Cellular localization of PELO-interacting proteins. A, HeLa cells were transiently co-transfected with PELO-HA and Myc-HAX1 and immunostained with anti-HA (panel i) and anti-Myc (panel ii) antibodies. A co-localization of both proteins is mainly diffused in cytoplasm (panel iii). B-N, Bimolecular fluorescence complementation analysis (BiFC) revealed that protein complexes between PELO and HAX1, EIF3G or SRPX are localized at the cytoskeleton. B-E, Cells transfected with GFPN (B) and GFPC (C), PELO-GFPC (D) and HAX1-GFPN (E) alone did not show any fluorescence. F-N, HeLa cells were transiently co-transfected with PELO-GFPC and either HAX1-GFPN (F), EIF3G-GFPN (G), SRPX-GFPN (M) or MOS-GFPN (N) as control. Complementation of both non-fluorescent GFP halves resulted in functional fluorophores, which is diagnostic for stable interaction of the fusion proteins. The reconstituted GFP-fluorescence (BiFC) is shown in green and mainly localized at the cytoskeleton. Absence of GFP-fluorescence (N) indicates lack of BiFC. DNA staining is shown in blue. Scale bars = 20 μm.