Fig. 6. Protein phosphatase 1 rapidly dephosphorylates Kalirin-P-T1590.
A. pEAK RAPID cells transiently transfected with vector encoding Kalirin7 were treated with phosphatase inhibitors for 30min before extraction in SDS lysis buffer. Duplicate aliquots of cell extract were visualized with antibody to P-T1590 (upper panel) or myc (lower panel). B. pEAK RAPID cells transiently transfected with parent vector or vectors encoding Kalirin7, ΔKalirin7 or KGEF1 were treated with or without 25 nM Calyculin A for 30min before extraction into SDS lysis buffer. Duplicates of cell extract were visualized with antibody to P-T1590 (upper panel) or myc (lower panel). C. PC12 cells expressing exogenous myc Kalirin7 and striatal cultures expressing endogenous Kalirin7 were exposed to medium containing 10μM roscovitine for 4h or 25nM Calyculin A for 30min (CA); control cells (Con) received vehicle.