Fig. 5

LPS treatment causes dynamin-dependent TLR-4 vesicular trafficking in WT BMMCs. The cells were pre-incubated for 30 min with either vehicle or dynasore (80 μM) and then stimulated with LPS (500 ng/mL). The cells were fixed and processed for confocal microscopy 15, 30, or 60 min after stimulus. a Representative photographs of cells showing TLR-4 (Red), Htt (green), and nuclei (blue). For reference, an ESID photograph is shown in each time point. Representative images are shown (n = 60–70 cells analyzed per condition). The images were taken at × 63 magnification. Scale bar = 2 μm. b, c Percentage of cells exhibiting perinuclear localization of TLR-4 and Htt, respectively. Pearson’s correlation coefficient for TLR-4 and Htt in d the whole area and in e the perinuclear area of the images obtained from the equatorial slices. Data are presented as the mean ± SD of at least three independent experiments, where 60–70 individual cells in three fields per preparation were analyzed. ^&p < 0.05, ^&&&p < 0.001 vs. the value at time zero or in non-stimulated cells; ###p < 0.001 vs. vehicle-treated cells as determined by two-way ANOVA and post hoc Tukey’s test