Vaccinia Virus--Induced Cell Motility Requires F11L-Mediated Inhibition of RhoA Signaling
Ferran Valderrama, João V. Cordeiro, Sibylle Schleich, Friedrich Frischknecht, Michael Way

Supporting Online Material

This supplement contains:
Materials and Methods
Figs. S1 to S6
Movies S1 to S4
References

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Movie S1.
This phase contrast movie shows a wound-healing assay of BS-C-1 cells in the absence of serum. The time-lapse covers a period of 15h in 6s, thus every second of the movie represents 2h 30min of real time. By following the cells during the course of the movie, it is evident that they are not able to invade the centre of the wound.

Movie S2.
This phase contrast movie shows a wound-healing assay of BS-C-1 cells infected with wild type Western Reserve (WR) strain of vaccina virus in the absence of serum. The time-lapse covers a period of 15h (from 5 to 20 hours post infection (hpi)) in 6s, thus every second of the movie represents 2h 30min of real time. By following the cells during the course of the movie, it is evident that WR infected BS-C-1cells are induced to migrate into the wound. Migrating cells are deficient in retraction of their rear and extend multiple lamellipodia at their leading edges.

Movie S3.
This phase contrast movie shows a wound-healing assay of BS-C-1 cells infected with the modified vaccinia Ankara (MVA) strain of vaccina virus in the absence of serum. The time-lapse covers a period of 15h (from 5hpi to 20hpi) in 6s, thus every second of the movie represents 2h 30min of real time. By following the cells during the course of the movie, it is evident that MVA infection does not stimulate BS-C-1 cells to become motile.

Movie S4.
This phase contrast movie shows a wound-healing assay of WR infected BS-C-1 cells previously transfected with siRNA against F11L in the absence of serum. The superimposed red channel reveals the signal from the control siRNA-3'-rhodamine GFP-22 oligo (Qiagen) that was co-transfected with the siRNA-F11L. This allows identification of infected cells in which F11L expression has been inhibited during the course of the movie. The time-lapse covers a period of 15h (from 5hpi to 20hpi) in 6s, thus every second of the movie represents 2h30min of real time. Infected cells in which there is a detectable rhodamine signal do not invade into the wound. A single non-transfected migrating cell is evident in lower right wound edge.

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