Rmm4534 028835

Lentiviral shRNA (ATG7 and BECLIN1) constructs were purchased from Open Biosystems (RMM4534_019584 and RMM4534_028835) (Ottawa, ON, Canada). ATG7 and BECLIN1 shRNAs were transfected into PC3, LNCaP, or MCF7 cells either alone or together with packaging plasmids following the manufacturer's protocol (Invitrogen ViraPower Lentiviral Expression Systems kit, Carlsbad, CA). Cells were passaged and plated in a six-well plate and allowed to adhere for 24 hours before infection. The silencing efficacy of the various shRNAs was assessed by Western blot analysis of ATG7 and BECLIN1 proteins using polyclonal antibodies (Sigma-Aldrich, Oakville, ON, Canada).

The PC3 cells with knockdown of ATG7 and BECLIN1 (or both) were generated. Lentiviral shRNA (ATG7 and BECLIN1) constructs were purchased from Open Biosystems (RMM4534_019584 and RMM4534_028835). The ATG7 and BECLIN1 shRNAs were transfected into PC3 cells, either alone or together with packaging plasmids following the manufacturer's protocol (Invitrogen ViraPower Lentiviral Expression Systems kit, Carlsbad, CA, USA). The silencing efficacy of the various shRNA was assessed by WB analysis of ATG7 and BECLIN1 proteins using polyclonal antibodies.