AMPK activation inhibits autophagosome maturation. (A) Formation rate and conversion ratio of LC3-positive and LC3-negative foci formed by Halo-ATG13, ULK1-Halo, Halo-WIPI2, and Halo-ATG2A in glucose-containing media in the absence and presence of the AMPK agonist MK8722. (B) Formation rate and conversion ratio of LC3-positive and LC3-negative foci formed by Halo-ATG13, ULK1-Halo, Halo-WIPI2, and Halo-ATG2A in media containing no glucose in the absence and presence of the AMPK inhibitor BAY3827. (C) Lifetimes of foci formed by Halo-ATG13, ULK1-Halo, Halo-WIPI2, and Halo-ATG2A before and during colocalization with GFP-LC3B in glucose-containing media in the presence and absence of the AMPK agonist MK8722 (left panels) or in media lacking glucose in in the absence and presence of the AMPK inhibitor BAY3827 (right panels). For all plots the box indicates the interquartile range, the whiskers the 10–90% confidence interval, the square indicates the average, and the horizontal line is the median; statistical significance was assessed by a two-tailed t-test.