Figure 8.

TP53INP2 silencing blocks beclin-1 recruitment to rapamycin-induced autophagic vacuoles. (a) HeLa cells were cotransfected with beclin 1-CFP, TP53INP2-EYFP, and LC3-ERFP plasmids and treated with rapamycin (10 μM for 4 h). TP53INP2 is in green, LC3 is in red, and beclin 1 is in blue. (b) HeLa cells were plated then, the next day, they were transfected with TP53INP2 siRNA or scrambled siRNA as a control and grown for an additional 24 h before beclin 1-CFP transfection. Twenty-four hours later, cells were treated with rapamycin (10 μM) for an additional 4-h period. (c) Relative quantification of beclin 1-CFP–positive vesicles in rapamycin-treated or untreated cells transfected with TP53INP2 siRNA or with scrambled siRNA as a control.