WISp39 binds phosphorylated Coronin 1B to regulate Arp2/3 localization and Cofilin-dependent motility

Files in this Data Supplement:

• Supplemental Material (PDF)
• Video 5 -
  Live-cell imaging of WISp39 KD cells without ZVAD. U2OS cells were transfected with WISp39 1B siRNA for 48 h and then imaged by time-lapse microscopy using an inverted microscope (IX81; Olympus). Frames were taken every 8 min for 24 h. Cells show an increase in bi- and multipolarity, chaotic migration and increased cell death compared with control cells. Bar, 50 µm.
• Video 6 -
  Live-cell imaging of WISp39 KD cells treated with ZVAD. U2OS cells were transfected with WISp39 1B siRNA for 48 h, treated with 100 µM ZVAD 1 h before imaging, and then imaged by time-lapse microscopy using an inverted microscope (IX81; Olympus). Frames were taken every 8 min for 24 h. Cells continue to show an increase in bi- and multipolarity and chaotic migration, but cell death decreases to control cell levels. Bar, 50 µm.
• Video 2 -
  Live-cell imaging of WISp39 KD cells. U2OS cells were transfected with WISp39 siRNA for 48 h and then imaged by time-lapse microscopy using an inverted microscope (TE-200; Nikon). Frames were taken every 7 min for 24 h. Cells show an increase in bi- and multipolarity with chaotic migration patterns unrelated to wound closure.
• Video 3 -
  Live-cell imaging of control cells without ZVAD. U2OS cells were transfected with control siRNA for 48 h then imaged by time-lapse microscopy using an inverted microscope (IX81; Olympus). Frames were taken every 8 min for 24 h. Cells show normal apolar morphology and directional migration similar to Video 1, with few dead cells. Bar, 50 µm.
• Video 1 -
  Live-cell imaging of control cells. U2OS cells were transfected with control siRNA for 48 h and then imaged by time-lapse microscopy using an inverted microscope (TE-200; Nikon). Frames were taken every 7 min for 24 h. Cells show normal apolar morphology and directional migration into the wound.
• Video 4 -
  Live-cell imaging of control cells treated with ZVAD. U2OS cells were transfected with control siRNA for 48 h, treated with 100 µM ZVAD 1 h before imaging, and then imaged by time-lapse microscopy using an inverted microscope (IX81; Olympus). Frames were taken every 8 min for 24 h. Cells showing normal apolar morphology and directional migration similar to Video 3, with few dead cells. Bar, 50 µm.
• Video 7 -
  Live-cell imaging of Coronin 1B siRNA-transfected cells. U2OS cells were transfected with Coronin 1B siRNA for 48 h and then imaged by time-lapse microscopy using an inverted microscope (TE-200; Nikon). Frames were taken every 7 min for 24 h. Cell morphology showing an increase in bi- and multipolarity is similar to WISp39 KD cells.