Figure 4.
Prazosin and Tamoxifen Lead to Lysosomal Leakage
(A) Analysis of whole cell proteome and cytosol fractions from MutuDCs treated with prazosin, tamoxifen, or DMSO (control) for 4 h. The relative abundance of proteins from prazosin or tamoxifen versus vehicle-treated cells in whole cell versus cytosol proteomes. The histograms show distributions of all (gray) versus lysosomal (purple) proteins. TM, transmembrane; SOL, soluble. p values were calculated using the Kolmogorov-Smirnov test. n = 2 for whole cell lysates (SILAC quantification), n = 4 for cytosol fractions (label-free quantification).
(B and C) MutuDCs stably expressing galectin-3-YFP were treated with 20 μΜ prazosin and imaged continuously for 40 min.
(B) Examples of control and prazosin-treated galectin-3-YFP-positive cells. Scale bar, 20 μm.
(C) Quantification of galectin-3-YFP recruitment in control and prazosin-treated cells from a representative movie. Each dot represents a sum of galectin-3-YFP spot areas per cell over the duration of the movie (40 min). Box and whiskers plot visualizes median, first, and third quartiles (hinges) and the smallest/largest observation no further than 1.5 * IQR (interquartile range) from the respective hinge (whiskers). p values were calculated using Mann-Whitney U test. Data representative of two experiments.
(D and E) MutuDCs were pulsed with 1 mg/mL 3K dextran-TRITC for 45 min and treated with 20 μΜ prazosin or tamoxifen for 1 h.
(D) Representative images. Scale bar, 20 μm.
(E) Quantification of the cytosolic fluorescence. Each dot represents a cell (three samples per condition). p values were calculated using Mann-Whitney U test with Bonferroni correction (comparing with the control group). Box and whiskers plot visualizes median, first, and third quartiles (hinges) and the smallest/largest observation no further than 1.5 * IQR from the respoective hinge (whiskers).