Fig. 2

FMNL2 expression induces filopodia formation. a C-terminally tagged full-length derivatives of each FMNL2 isoform were expressed in NIH 3T3 fibroblasts by transient transfection. F-actin (green) was detected with phalloidin and FMNL2 expression (red) was detected by virtue of the C-terminal mCherry tag. Nuclei were detected with DAPI (blue) in merged image. FMNL2 expression induced extensive filopodia formation in comparison to untransfected cells or cells expressing mCherryFP with an N-terminal myristoylation motif (top panel). b N-terminally tagged full-length derivatives of each of the FMNL2 isoforms were expressed in NIH 3T3 cells by transient transfection. F-actin (green) was detected with phalloidin and FMNL2 expression (red) was detected by virtue of the N-terminal myc tag. Expression of N-terminally tagged FMNL2 isoforms does not induce filopodia formation. mCherryFP was included as a control (top panel) c Quantification of data shown in (a, b). “C” indicates C-terminal mCherry tag, “N” indicates N-terminal myc tag. N = 3, >100 cells counted per sample, error bars = SEM. d NIH 3T3 fibroblasts were transiently transfected with an SRF luciferase reporter gene and the indicated full-length FMNL2 derivative. “C” indicates C-terminal mCherry tag, “N” indicates N-terminal myc tag. Reporter activation is expressed relative to an SRF-VP16 control fusion protein. N = 3, error bars = SEM. e Equivalent samples of transfected cell lysates were subjected to SDS-PAGE and immunoblotted. Expression of the indicated FMNL2 isoforms was detected with either anti-mCherry or anti-myc antibody