Analysis Tools
cellular
|
• phalloidin-stained mutant MEFs display an altered shape with broader and more stable protrusions, as determined by their reduced cell body solidity (cell body outline divided by convex hull) relative to wild-type MEFs
• as early as 1 min after PDGF stimulation, mutant MEFs show increased formation of circular dorsal ruffles (CDRs) relative to wild-type MEFs
• mutant MEFs require lower doses of PDGF to initiate CDRs relative to wild-type MEFs
|
|
• mutant MEFs exhibit delayed focal adhesion (FA) disassembly relative to wild-type cells; at 10 min after nocodazole washout, mutant MEFs still retain ~90% of vinculin-positive FAs, unlike in wild-type MEFs where ~50% of FAs are already lost
• at steady state, mutant MEFs display an increased number of smaller FAs with reduced paxillin intensity relative to wild-type MEFs, suggesting impaired FA maturation
|
|
• mutant MEFs migrate with significantly increased directionality, unlike wild-type MEFs which frequently change direction following a meandering path
• mutant MEFs display increased maximum trailing end length before rear detachment and increased trailing end lifetime relative to wild-type MEFs
• however, no consistent change in random migration velocity is observed
|
|
• mutant MEFs show a striking accumulation of the proteoglycan CSPG4 (NG2) on the cell surface due to impaired internalization relative to wild-type MEFs
• mutant MEFs display CSPG4 (NG2) clusters constitutively, whereas wild-type MEFs only upon stimulation with PDGF or upon inhibition of endocytosis with dynasore
|
