Fig. 5
From: Tight association of autophagy and cell cycle in leukemia cells
Cyto-ID-sorted cells display different autophagy levels over at least 24 h. Cells were flow-cytometrically sorted on the basis of their Cyto-ID fluorescence intensity into subpopulations with low, medium, and high Cyto-ID fluorescence (AutLO, AutME and AutHI, respectively). A Representative immunoblots of lysates from sorted cells. Lysates were prepared after 1-h cultivation of sorted cells in the absence or presence of 10 µM CQ. B Sorted cells were incubated for the indicated times, and autophagy was determined by flow-cytometric analysis of Cyto-ID-stained cells. Cyto-ID fluorescence intensities of the three fractions were normalized to the Cyto-ID fluorescence intensity of AutME. C, D RNA was prepared either approximately 1 h after sorting (C) or after 24-h cultivation of sorted cells (D). mRNA expression levels were determined by real-time RT-PCR and normalized to B2M expression levels. Mean ± SEM of three (C–D) or four (B) independent measurements is shown (*P < 0.05)