Figure 2
From: A large-scale RNA interference screen identifies genes that regulate autophagy at different stages
A large-scale RNA interference screen identifies autophagy-regulating genes. (A) Schematic diagram illustrating a large-scale RNA interference screen. 336 shRNAs were identified in MDC positive cells. (B) Candidate validation using the Cyto-ID spectrophotometric assay. K562 cells were treated with non-silencing (NS) shRNA or individual 336 candidate shRNAs. The 1.4-fold increase of Cyto-ID fluorescence was empirically set as the cut-off line. (C) Candidate validation using the LC3B immunoblotting assay. K562 cells were treated with NS or 152 individual shRNAs of Cyto-ID positive candidates. Autophagy was monitored using LC3B immunoblotting. A cropped representative blot of LC3B immunoblotting is shown. Full images are included in supplemental materials. (D) Quantification of LC3B-II intensities. The intensities of protein bands were quantified using Image J. The ratios of the protein intensities of LC3B-II and ACTB are shown. The 1.4-fold increase of LC3B-II was empirically set as the cut-off line.
