Fig. 4

Beclin-1 knockdown does not block autophagy induction in ovarian cancer cell lines with low, intermediate, and high levels of Beclin-1 expression. CaOV3, OVCAR8 eGFP-LC3, and HeyA8 cells were each transfected with control siRNA (siNT) or siRNA targeting BECN1, ATG5, ATG7, or ATG5 + 7 and cells seeded to adherent or non-adherent culture. a Adherent cells were allowed to attach overnight, treated with DMSO or Akti-1/2 (5 μM) the next day, and harvested 24 h later to generate protein lysates. Immunoblots were performed for indicated proteins. b Spheroids (along with a parallel adherent culture transfected with control siRNA) were harvested 24 h after seeding to non-adherent culture. Protein lysates were generated and immunoblot performed for indicated proteins. Immunoblots depicted are representative of duplicate experiments