strong kinetic coupling between cortical actin flow and inward TCR MC movement was maintained after each drug treatment. This strong coupling is in contrast to a previous report using bilayer engaged Jurkat T cells, in which the rate of inward TCR natural product library MC movement at the periphery of the IS was reported to be ?40% the rate of centripetal actin flow. As described in more detail in the Introduction, this and other studies?especially those that have characterized the effects of physical barriers within the bilayer on the rates of TCR MC movement ?have led to a dissipative or frictional coupling model of TCR MC actin cytoskeleton interaction that allows slippage between the MC and actin flow. Although we certainly believe that such slippage would occur if we had used physical barriers, we think that in the absence of such barriers the coupling between TCR MCs and actin flow is probably quite tight.

That said, at least part of the difference Chromoblastomycosis between our study and that of Kaizuka and colleagues as regards the kinetic coupling between actin flow and TCR MC movement could be due to possible differences in bilayer conditions between the two studies. It is also possible that, in the study by Kaizuka et al., the quantitation of actin flow rate was restricted largely to the LP/dSMAC, whereas the quantitation of TCR MC movements was made predominately in the LM/pSMAC, leading to the discrepancy between their respective centripetal rates. Obviously, much remains to be learned regarding the components and physical properties of the mechanisms that couple TCR MCs and integrin clusters to cortical actin flow during IS formation. Our demonstration that TCR MCs exhibit two distinct rates of centripetal movement across the IS can actually be reconciled with a large number of rates reported previously.

First, the fast rate across the LP/dSMAC Cathepsin Inhibitor 1 reported here corresponds relatively well with rates at the periphery of the IS reported by several groups. Moreover, the slow rate across the LM/pSMAC reported here corresponds quite well with rates reported for regions of the IS that are almost certainly inside the LP/dSMAC, that is, to the LM/pSMAC. Thus our finding that TCR MCs move at different speeds depending on the region of movement, that is, the LP/ dSMAC versus the LM/pSMAC, helps to reconcile the wide range of speeds reported previously for TCR MC movements at the IS. The role of myosin IIA at the IS As discussed in the Introduction, the role of myosin IIA in IS formation has been somewhat controversial.

Specifically, an earlier study using BB argued that myosin IIA is not required for IS formation, whereas a more recent report using BB and RNAi mediated knockdown of myosin II argued that the myosin is required for significant TCR MC transport, cSMAC formation, and IS stability.