Casein Kinase Iγ2 Impairs Fibroblasts Actin Stress Fibers Formation and Delays Cell Cycle Progression in G1
Figure 3
The catalytic activity of CKIγ2 is required to induce change in cell morphology and actin stress fibers in fibroblasts. (a) Isolated clones of fibroblasts stably transfected with a plasmid encoding HA-tagged wild-type CKIγ2 (Z23), HA-tagged kinase deficient CKIγ2 (KD1 and KD30), or pool of stable fibroblasts transfected with an empty plasmid (R2Zeo) were analyzed for HA-CKIγ2 expression by HA Western Blot (WB) on HA immunoprecipitates (IP) and CKIγ2 activity as determined on HA immunoprecipitates from equal amounts of protein normalized cell lysates and in vitro kinase assays using α-casein as substrate. KD1 and KD30: clones that overexpress kinase deficient CKIγ2 at the same levels as cells stably overexpressing wild-type CKIγ2 (Z23). (b) Morphology of serum growing cells was visualized by phase contrast microscopy (40X) (upper panels) and actin organization by actin staining with rhodamine-conjugated phalloidin (63X) (lower panels).
