Fig. 5

The cholesterol depletor MβCD modulates the uptake and subcellular localization of RGD-C’Dots in osteocytes in vivo. A-D) Short incubation experiments highlight changes between non-treated and MβCD treated groups over time via linear regressions of cell count and t-tests of subcellular localization. There were no differences in slope, but there were significant differences in the y-intercept between short incubation groups (differences indicated by # symbols). E-H) Long incubation experiments highlight how cholesterol depletion causes an increase in osteocyte uptake and retention of C’Dot signal. 2-Way ANOVA’s compare the number of cells with C’Dot signal between groups and t-tests compare the percentage of cells with subcellular localization of C’Dot signal. In these long incubation experiments, both sexes display an increase in C’Dot retention with MβCD application. Differences in ANOVA post-hoc tests between long incubation groups are shown at each timepoint with * symbols. The response of other C’Dot surface functionalized groups to MβCD preincubation is shown in Supplemental Figs. 4 and 5. I) Cholesterol and lipid raft mediated endocytosis is broadly active uptake pathway, regardless of nanoparticle functionalization, and is disrupted with application of MβCD. (*p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001; Error bars indicate SEM).