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. 2019 Jan 17;9:151. doi: 10.1038/s41598-018-36379-z

Figure 2.

Figure 2

More autolysosomes are present in CLN5-deficient patient fibroblasts. (A) Control or CLN5-deficient fibroblasts that had been transfected with the mRFP-GFP-LC3 plasmid for 24 hours were fixed. GFP and RFP signal were visualized using confocal microscopy. The fraction of RFP/GFP signals that was overlapping was analyzed by the Mander’s coefficient using the ImageJ JACoP plugin. In ImageJ, two channels of each image file were first split to two separate images. These two images were then used for plugin JACoP analysis. The threshold of each image was adjusted to reduce the background signals before performing JACoP Mander’s coefficient. n = 15 cells, error bar represents SEM. (B) Control fibroblasts that had been transfected with the mRFP-GFP-LC3 plasmid for 24 hours were treated with CQ, baf A, or HBSS for 2 h prior to fixation. The fraction of RFP/GFP signals that was overlapping was analyzed by the Mander’s coefficient using the ImageJ JACoP plugin. n = 7 cells, error bar represents SEM. All experiments were repeated at least three times. Scale bar: 20 μm.