Fig. 7: Temporospatial features of endocytosis enhancement after challenge with recombinant PLY.
a Time frame of endocytosis change (FM4-64, normalized to timepoint 0, n = 6 independent experiments), exosome release (CellMask, single vesicles counted, n = 4 independent experiments), membrane depolarization (DiBac4(3), fluorescence increase, normalized to timepoint 0 and to mock, n = 15 cells), calcium increase (Fura-2, ratiometric value, increasing with calcium increase, n = 4 independent experiments), and cell swelling (calcein AM, fluorescence intensity inversely normalized to timepoint 0 (the value increases when swelling increases, but in reality fluorescence falls)). b Scheme of the hanging basket with a 1 µm porous membrane allowing localized treatment through the pores of the underside of the cells (incubation with 2 HU/ml PLY), prestained with FM 4-64. c FM 4-64 fluorescence increase along the membrane with pores (red curve), normalized to the fluorescence along the top membrane (blue curve), demonstrates the localized nature of the toxin’s pro-endocytotic effect. Values represent the mean ± SEM. n is indicated and represents single cells tested, pooled together from three independent experiments. Source data are provided as a Source Data file.
