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Fig. 6. The role of F-actin in Shrm-induced constriction. (A-D) Cells expressing ShrmL (A and B) or Endo-ShrmC cells (C and D) were treated with DMSO (A and C) or 1 µM cytochalasin D (B and D) for 30 minutes and stained to detect ZO-1. (E,F) Wild-type T23 cells grown on Transwell filters were treated with DMSO (E' and E'') or 2 µM CD (F' and F'') for 30 minutes, stained with phalloidin to detect F-actin and imaged by confocal microscopy to visualize the cytoskeleton at either the apical or basal surface. (G-I) Control (G), ShrmL expressing (H), or Endo-ShrmC expressing (I) cells were treated with 2 µM cytochalasin D for 30 minutes and stained to detect E-cadherin (green), ZO-1 (red, H' and I'), or Shrm (blue, H'' and I''). (J-L) Control (J), ShrmL expressing (K), or Endo-ShrmC expressing (L) cells were pre-treated with 100 µM blebbistatin for 30 minutes followed by 2 µM cytochalasin D for 30 minutes and stained to detect E-cadherin (green), ZO-1 (red, K' and L'), or Shrm (blue, K'' and L''). Scale bar represents 15 µm in all panels.
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