Figure 2

Normal human mammary cell subsets contain different levels of ATG proteins and autophagosomes. (A) Representative FACS profiles of human mammary subsets isolated from three normal donors and WBs of their content of different autophagy proteins with H3 as an internal loading control. (B) Scatter plots showing the corresponding quantification of human LC3B-II, ATG7 and BECLIN1 normalized to H3 measured independently in subsets isolated from the same three normal samples shown in (A). (C) Effect of a two-hour exposure of FACS-purified BCs and LPs from three different normal donors (A) to 200 nM BafA₁ (Baf [ +]) or an equivalent concentration of DMSO (0.5%) on their content of LC3B-II (relative to ACTIN). (D) Distribution of R⁺ and G⁺ cells in FACS profiles of RFP-GFP-LC3B-transduced BCs and LPs analyzed after four days in culture. (E) Levels of ATG7, P62, ATG4B, ACTIN and LC3B determined in WBs of the G⁺ and R⁺ cells isolated from the four-day cultures shown in (D). Bar graphs show the ratios of LC3B-II/LC3B-I and P62/actin levels in the four subsets of cells analyzed. P-values were calculated using a paired t-test. *P < 0.05, **P < 0.01.