Cell motility is restricted by membrane targeting of PKCδ.

<p>a). Live cell images of stably transfected PKCδ-CaaX and PKCδ-SaaX NR6-WT (EGFR) cells with endogenous knockdown of PKCδ were taken every 30 minutes with and without 1 nM of EGF in quiescent media for 24 hour time period. Cell motility was analyzed by metamorph software. (Two-Sample-t-test was utilized to evaluate significance p<.05, n>11). b) Live cell images of stably transfected PKCδ-CaaX and PKCδ-SaaX NR6-WT (EGFR) cells with endogenous knockdown of PKCδ were taken every 15 minutes stimulated with 5nM of EGF and 5 nM of insulin for 5 hours. Average individual cell speed/path was analyzed at 15 minute intervals for a total duration of 5 hours with metamorph software. (Two-Sample-T-test was utilized to evaluate significance (p < 0.05, n >11). c) Stably transfected PKCδ-CaaX and PKCδ-SaaX NR6-WT (EGFR) were grown in a 6 well plate to 90% confluency prior to scratching with rubber policeman. Cells were treated with 1 nM of EGF in quiescent media and images of scratch were taken at 4X objective magnification. d,e,f,g) Stably transfected PKCδ-CaaX and PKCδ-SaaX NR6-WT (EGFR) <i>cells </i><i>were </i><i>grown </i><i>to 90% confluency prior to plating on collagen I coated plate. Cells were then allowed to adhere overnight and then transfected with 50uM of</i> PKCδ specific siRNA and stimulated with 1 nM EGF. Cells were then imaged for 12 hours with 30 minute intervals and average individual cell speed (d) /path (e) /persistence (absolute displacement from origin)(f) was analyzed with metamorph software. (g) Persistence was analyzed utilizing the median of absolute angle standard deviation relative to control.