Fig. 3.

Efficient cytosolic trafficking of all CPMPs and CPPs requires the human HOPS complex. (A) Figure legend. (B–F) The effect of VPS39 and VPS41 knockdown (HOPS-specific subunits) compared with TGFBRAP1 and VPS8 knockdown (CORVET-specific subunits) on both overall uptake (FC) and cytosolic access (FCS) of 1^R (B), 2^R (C), 3^R (D), 4^R (E), and 5^R (F) relative to the effect of nontargeting siRNA (NT siRNA). FC: total cell uptake, fluorescence intensity at 585 nm in arbitrary units (a.u.). n refers to the number of biological replicates. For each FC replicate, the median fluorescence intensity at 585 nm was measured for at least 10,000 Saos-2 cells (gated for live cells). FCS: cytosolic and nuclear concentration (nM) (B–D) or intensity (MHz) (E and F). Each data point (n) denotes one 50-s FCS measurement recorded in the nucleus or cytosol of a single Saos-2 cell with (B–D) or without (E and F) autocorrelation. Error bars represent the SEM. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05, not significant (ns) for P > 0.05 from one-way ANOVA with Dunnett post hoc test.