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Fig. 5. The mutant Rab7T22N induces the accumulation of LC3-labeled autophagosomes. (A) CHO cells expressing EGFP-Rab7 wt or the mutant T22N were transiently transfected with the plasmid pCI-neo Myc-LC3. After 36 hours transfection, cells were subjected to indirect immunofluorescence as described under Materials and Methods and analyzed by confocal fluorescence microscopy. Briefly, cells were fixed, permeabilized and incubated with an antibody against the myc epitope. The corresponding Texas red-conjugated secondary antibody was used. Cells were mounted with Mowiol. (B,C) The number (B) and size (C) of LC3-labeled vacuoles per cell were quantified by the Methamorph software, using the integrated morphometry analysis. Data represent the mean±s.e.m. (n=25 cells). **P<0.01
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