Extended Data Fig. 1: Localisation of NPFs in blebbing melanoma cells.
From: SPIN90 associates with mDia1 and the Arp2/3 complex to regulate cortical actin organization
(a-d) Confocal microscopy images of blebbing M2 melanoma cells expressing GFP-tagged NPFs. White arrowheads indicate expanding blebs and red arrowheads retracting ones. Scale bars=5μm. Experiments were repeated two times independently with similar results. a, IQGAP1 localizes to the rim of retracting blebs (red arrowhead) but not to the rim of nascent blebs (white arrowhead). b, SPIN90-GFP before (left) and after (right) permeabilization and fixation to remove cytoplasmic signal. Following permeabilization and fixation, SPIN90 remains localized to the actin cortex of retracting blebs but is absent from the cortex of nascent blebs. c, FLII-GFP before (left) and after (right) permeabilization and fixation. Following permeabilization and fixation, FLII remains localized to the actin cortex of retracting blebs. d, Constitutively active cortactin-GFP localizes to the rim of retracting blebs but not to the rim of nascent blebs.
