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The Mid1-binding domain of Alpha4 is essential to protect PP2Ac from degradation. HEK293FT cells were transfected with HA3-PP2Ac alone or with HA3-PP2Ac and FLAG-Alpha4, FLAG-Alpha4ΔC, or FLAG-Alpha4_ED. Cells were treated with 50 μm cycloheximide (CHX) 48 h post-transfection and then lysed at the indicated time points post-treatment. The lysates were subjected to Western analysis using antibodies recognizing Alpha4, PP2Ac, and HSP90.
