Figure 5.
Figure 5. HRS is required for MT1-MMP endosomal recycling. (A) MDA–MB-231 cells were treated twice with siRNA over 120 h before fixation and subsequent treatment with guanidinium hydrochloride. Sum intensity projections. Bars: (main images) 10 µm; (insets) 2 µm. MRG, merge. Brightness and contrast of main images and insets was adjusted uniformly across panels to best display plasma membrane or endosomal staining, respectively. (B) Quantification of sum intensity of MT1-MMP in a VPS35 mask. n = 50 cells over three independent experiments. *, P < 0.05. Statistical analysis, one-way ANOVA, Dunnett’s post hoc test. (C) MDA–MB-231 cells were treated with siRNA targeting HRS for 120 h and transfected with paCherry–MT1-MMP and GFP-EEA1. Trafficking from the endosome was measured by quantifying the decrease in paCherry fluorescence in the endosome. n = 6 independent experiments; >20 cells total; error bars indicate SEM.

HRS is required for MT1-MMP endosomal recycling. (A) MDA–MB-231 cells were treated twice with siRNA over 120 h before fixation and subsequent treatment with guanidinium hydrochloride. Sum intensity projections. Bars: (main images) 10 µm; (insets) 2 µm. MRG, merge. Brightness and contrast of main images and insets was adjusted uniformly across panels to best display plasma membrane or endosomal staining, respectively. (B) Quantification of sum intensity of MT1-MMP in a VPS35 mask. n = 50 cells over three independent experiments. *, P < 0.05. Statistical analysis, one-way ANOVA, Dunnett’s post hoc test. (C) MDA–MB-231 cells were treated with siRNA targeting HRS for 120 h and transfected with paCherry–MT1-MMP and GFP-EEA1. Trafficking from the endosome was measured by quantifying the decrease in paCherry fluorescence in the endosome. n = 6 independent experiments; >20 cells total; error bars indicate SEM.

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