FIGURE 2:

Cbl and Cbl-b facilitate CSF-1R internalization and translocation to the macropinosome lumen. (A) Immunofluorescence of cell surface CSF-1R in WT and DKO macrophages at indicated CSF-1 stimulation period. (B) Quantification of surface CSF-1R at different time points, scale bars are SD with n = 100, same letters are not significantly different (p< 0.05) by 2-way ANOVA followed by Tukey HSD comparison of means. (C) Phase contrast images illustrating the number of macropinosomes in DKO BMDM. (D) Quantified number of macropinosomes per cell (error bars = std. dev., n = 100 macrophages across two experiments). (E) Delivery of the CSF-1R to the macropinosome visualized labeling by sequential imaging of Texas-red dextran (macropinosome) and CSF-1R immunofluorescence following permeabilization (scale bar = 5 μm). (F) Quantification of the percentage of macropinosomes that contain luminal CSF-1R (error bar = std. dev. with n = 50 from two independent experiments). (G) CSF-1Rimmunofluorescence delivery to macropinosomes and degradation (scale bar = 10 μm). (H). Colocalization of CSF-1R and Hrs 10 min after CSF-1 exposure (scale bar = 10 μm).