Figure 3

Characterisation of truncated YFP-Septin2 constructs. (A) Schematic presentation of full length YFP-Septin2 (YFP-Sept2-PB/G/CC) and truncated YFP-Septin2 constructs. (B) In some cells overexpression of full length YFP-Sept2-PB/G/CC induced the formation of higher-ordered YFP-Septin2 containing-structures with differing morphology (C) Untagged full length Sept2-PB/G/CC is also capable of forming these structures and actin filaments are attenuated in those cells. Full length Sept2-PB/G/CC was detected with anti-Septin2-antibody. (D) Distribution of YFP-Septin2 constructs in NRK cells and their effect on the actin cytoskeleton. Cells were transfected with either full length YFP-Sept2-PB/G/CC or the truncated YFP-Septin2 constructs and stained for actin with phallacidin. Only YFP-Sept2-PB, the construct lacking the GTPase domain and the coiled coil domain affects actin organisation. The distributions of YFP-Sept2-PB/G, the construct without the coiled-coil domain, and YFP-Sept2-G/CC, which lacks the polybasic region, are different to full length YFP-Sept2-PB/G/CC, but there is no effect on actin. Images show a single confocal section. Bars. 10 μm.