Extended Data Fig. 4: Transfection of Effectene-coated beads induces TFEB nuclear localization in an ATG conjugation system-dependent manner.
(a) Representative immunofluorescence images showing TFEB::mNeonGreen and Gal-3 counterstained by DAPI. Transfection of Effectene-coated beads damaged endosomes or lysosomes, whereas transfection with PEI-coated beads did not. Note that cells harbouring the damage visualized by Gal-3 exhibited TFEB nuclear translocation (open arrowheads). DIC images showed that transfected beads were internalized in cells. (b) Ratio of nuclear to cytoplasmic TFEB::mNeonGreen in WT and ATG7-KO cells transfected with either Effectene- or PEI-coated beads. More than 100 cells were analysed per condition by cell profiler and the experiments were repeated three times. Bars represent mean ± s.d. (n = 3 biologically independent experiments, one-way ANOVA with Tukey’s test, **P = 0.001 for WT PEI versus effectene, P = 0.9711 for ATG7-KO PEI versus effectene) Scale bars, 50 μm. Statistical source data are provided in Source Data Extended Data Fig. 4.
