Fig. 3
From: PTEN is a negative regulator of mitotic checkpoint complex during the cell cycle
PTEN suppresses MCC formation during the cell cycle. HeLa cells co-transfected with siRNAs to PTEN (siPTEN), or to luciferase (siLuc) as control, and Flag-Mad2 expression plasmid were synchronized at G1/S junction via double-thymidine treatment. At various times after releasing into the cell cycle, cells were collected and lysed. Equal amounts of cell lysates were immunoprecipitated with the anti-Flag antibody. Flag immunoprecipitates, along with lysate inputs, were blotted for Flag, PTEN, Cdc20, BubR1 and Bub3. TT release double thymidine release