Figure 7.

A role for autophagy in IKBKE-dependent normal breast epithelial cell transformation and TNBC proliferation. (a) Proliferation assay of MDA-MB-231 cells upon downregulation of endogenous IKBKE, ATG5, ULK1 protein levels by transfection with the indicated siRNA (scrambled siRNA as negative controls, and unrelated specific siRNA against human IKBKE, #679, #680 and siRNA for ATG5 and ULK1). Cell viability was evaluated 72 h post-transfection by counting cells in triplicate with a Z2 Coulter Counter. Data were processed in Prism 6 software. Results from one experiment, representative of 3 independent experiments (n = 3) are shown. (b) Same as in (a), but using MDA-MB-468 TNBC cells. (c) Cell count of 1-7HB2 cell stably expressing IKBKE (WT), evaluating cell proliferation, by counting cells in triplicate with a Z2 Coulter Counter, upon downregulation (72 h post-transfection) of endogenous ATG5 and ULK1 with specific siRNA. A scrambled siRNA was used as a negative control. Data were processed with Prism 6 software. Results from one experiment, representative of 3 independent experiments (n = 3) are shown. (d) Proliferation assay of 1-7HB2 cell stably expressing IKBKE (WT) upon pharmacological inhibition of autophagic activity by SAR-405 (10 μM) and Spautin-1 (100 μM). Cell proliferation was evaluated after 72-h treatment by counting cells in triplicate with a Z2 Coulter Counter. Data were processed with Prism 6 software. Results from one experiment, representative of 3 independent experiments (n = 3) are shown.