Optogenetic actin network assembly on lipid bilayer uncovers the network density-dependent functions of actin-binding proteins
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Finally, we confirmed that this actin polymerization is mediated by the Arp2/3 complex; the cells treated with CK-666, an Arp2/3 complex inhibitor, showed a relatively weaker increase in the cortical Lifeact signal compared to the DMSO-treated control cells during illumination (Fig. 1f, 1g), although the translocation dynamics of SspB-mScarlet-I-VCA were nearly identical between the two conditions
I found this result a bit puzzling with respect to the maintenance of a strong cortical network in the presence of CK-666 (which could be due to timing of the experiment if there was insufficient time for branched network turnover and inhibition of newly nucleated actin branches) but then began to wonder if part of the issue is due to this being done in trypsinized cells shortly after replating -- I'm sure I'm not thinking of something …
Finally, we confirmed that this actin polymerization is mediated by the Arp2/3 complex; the cells treated with CK-666, an Arp2/3 complex inhibitor, showed a relatively weaker increase in the cortical Lifeact signal compared to the DMSO-treated control cells during illumination (Fig. 1f, 1g), although the translocation dynamics of SspB-mScarlet-I-VCA were nearly identical between the two conditions
I found this result a bit puzzling with respect to the maintenance of a strong cortical network in the presence of CK-666 (which could be due to timing of the experiment if there was insufficient time for branched network turnover and inhibition of newly nucleated actin branches) but then began to wonder if part of the issue is due to this being done in trypsinized cells shortly after replating -- I'm sure I'm not thinking of something obvious but why not just grow and image directly on chamber slides to preserve the cortical actin network in a more native state?
Regardless, I also wanted to mention that I found this to be an very elegant study using an approach that indeed can illuminate some important properties of actin binding protein behavior.
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