Figure 1

Determination of saturating concentrations of BafA and CQ by Western blotting in HeLa cells. (A,B) Treatment of HeLa cells for 24 h with various concentrations between 1.25 and 80 nM BafA or 0.1–300 µM CQ. Top: representative Western blots for LC3, p62/SQSTM1 and actin; bottom: quantifications from 3 independent experiments (mean ± SEM). For quantification, LC3-II levels at each concentration were normalized to the LC3-II levels at the highest concentration applied. (A) BafA increased LC3-II levels with an EC₅₀ of 5.6 nM and p62/SQSTM1 levels with an EC₅₀ of 6.5 nM. (B) For CQ, the calculated EC₅₀ for LC3-II and p62/SQSTM1 increase were 30 µM and 7.2 µM, respectively. For curve fitting, a non-linear regression curve fit (log(inhibitor) vs. response- variable slope (four parameters)) was applied.