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. 2011 Feb 15;22(4):467–477. doi: 10.1091/mbc.E10-02-0129

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© 2011 Tan et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 4: — RNAi of LIC1 and LIC2 causes enlarged and brighter lysosomes and disrupts lysosomal distribution but has no effect on early endosomes or the Golgi. (A) Rat2 cells were treated with scrambled, LIC1, or LIC2 RNAi and then costained for LAMP1 and LIC1 or LIC2. (B) Rat2 cells transfected with LIC siRNA and siRNA-resistant HA-LIC1 or Flag-LIC2 rescue constructs and costained for LAMP1, HA, or Flag and for DNA with DAPI. Gray lines delineate individual cell boundaries of siRNA-resistant LIC-overexpressing cells. (C) Quantification of the percentage of cells displaying altered late endosome/lysosome phenotype (left graph) and for the percent cell area of LAMP1 fluorescence per cell (right graph). The total area of positive LAMP1 staining within a cell is shown as a percentage of total cell area. Averages are shown where at least 30 cells were counted per condition from each of three experiments. (D, E) Rat2 cells were cotransfected with GFP and either scrambled RNAi or both LIC1 and LIC2 RNAi together and stained for Golgi with anti-GM130 (D) or early endosomes with anti-EEA1 (E) in red and for DNA with DAPI in blue. Gray lines delineate individual cell boundaries of transfected cells. Scale bars, 5 μm.