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. Author manuscript; available in PMC: 2019 May 3.
Published in final edited form as: Cell. 2018 Mar 22;173(4):851–863.e16. doi: 10.1016/j.cell.2018.03.010

Figure 4. Cold-induced Lysosomal Membrane Permeabilization (LMP) in Human iPSC-neurons.

Figure 4

(A) (Left panel) Live imaging of lysosomes in GS and human iPSC-neurons with DND-26 (green) and Magic Red (magenta; see STAR METHODS). Note: Following 1-h incubation at 4°C, accumulation of diffuse cytoplasmic fluorescence in marked human neurons (arrowheads) indicating LMP. Scale bar: 20 μm. (Right Panel) Quantification of cells containing diffused Magic Red signals (Diameter > 5μm) (n = 6 experiments; Student’s t-test for two-group comparison; *** p < 0.001).

(B) (Left panel) Live imaging of lysosomes in human iPSC-neurons with Magic Red under denoted conditions. Note: BAM15 (0.1 μM) or the antioxidant vitamin C (0.5 mM) alleviated LMP in human neurons treated following 4-h incubation at 4°C. Scale bar: 20 μm. (Right Panel) Quantification of cells containing diffused Magic Red signals (Diameter > 5μm) (n = 5 experiments; Student’s t-test between untreated controls with or without cold exposure; ANOVA plus post hoc Tukey test for multiple cold-exposed groups; ** p < 0.01; *** p < 0.001). See also Figure S5 for the involvement of other PQC components. For these experiments, neurons were derived from 2 GS and 3 human iPSC lines for each group.