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. 2016 Apr 11;213(1):81–95. doi: 10.1083/jcb.201510002

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© 2016 Hashimoto et al.

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Figure 1. — TGFβ1 activates Arf6 to promote the invasion of MDA-MB-231 cells. (A) Arf6 activities upon the stimulation of serum-starved cells by TGFβ1 for the indicated times were measured by detecting Arf6-GTP and total Arf6 by anti-Arf6 immunoblotting. (B) Matrigel invasion activities were measured in cells pretreated with the indicated siRNAs or siRNA with an irrelevant sequence (Irr) in the presence or absence of TGFβ1 or HGF. The number of cells that invaded through the Matrigel to the lower surface of the filters was scored as described in the Matrigel invasion assay section within Materials and methods. (C) TGFβ1- and HGF-induced Arf6 activation were measured in cells pretreated with siRNAs for GEP100 or Irr. (D) Activation of c-Met upon TGFβ1 stimulation was detected by a c-Met pY1234/1235 antibody together with total c-Met by immunoblotting. (E and F) TGFβ1-induced Arf6 activities (E) and Matrigel invasion activities (F) were measured in cells pretreated with the indicated siRNAs or Irr. Matrigel invasion activities were measured as in B. The results represent mean ± SEM. n = 3. *, P < 0.001. Total, total cell lysate.