Figure 2

A. Doxycycline has no effect on PP1α protein expression in control cell lines. Equal quantities (50 ug) of whole-cell lysates harvested in the absence (indicated above the lanes with a (-) symbol), or presence (at 24 hr following addition, indicated above the lanes with a (+) symbol), of doxycycline were separated by SDS-PAGE and subjected to immunoblotting using antibody specific for PP1α. UMUC3 – parent cell line from which stable expressor LLWO2F was selected; LLWO1 – cell line generated from UMUC3 which was stably transfected with the reverse tetracycline transactivator plasmid pUHD172-1neo only. B – Doxycycline induces a protein in LLW02F cells that is recognized by antibody to hemagglutinin. Equal quantities (50 ug) of whole-cell lysates harvested at the various time points in hours after doxycycline addition (indicated above each lane) were separated by SDS-PAGE and subjected to immunoblotting as described in Materials and Methods. C – Time-course of recovery from doxycycline induction. LLWO2F cells were induced for 24 hr. At the end of this time, equal quantities (50 ug) of whole-cell lysates harvested at the various time points in hours following removal of doxycycline (indicated above each lane) were separated by SDS-PAGE and subjected to immunoblotting with antibody to hemagglutinin.