Skip to main content
. Author manuscript; available in PMC: 2016 Dec 3.
Published in final edited form as: Cell Rep. 2016 Nov 15;17(8):1950–1961. doi: 10.1016/j.celrep.2016.10.062

Figure 4. Dynein-mediated centripetal transport of lysosomes is independent of BORC.

Figure 4

(A) A dominant-negative mutant of the dynactin p150 subunit (mCh-p150-CC1) was expressed in WT, myrlysin-KO and diaskedin-KO HeLa cells to inactivate dynein-mediated retrograde transport of lysosomes. Cells were stained with antibody to endogenous LAMTOR4 and analyzed by immunofluorescence microscopy. (B) Schematic representation of the constructs designed for accumulation of lysosomes at the cell periphery and their release after treatment with biotin. (C,D) Fluorescence microscopy and quantification of lysosome distribution (performed as explained in the legend to Figure 1D) showing changes in lysosome positioning after biotin-induced release from their peripheral sites in (C) WT and (D) myrlysin-KO HeLa cells. Student's t-tests (at 15 and 30 min of treatment vs. initial time point) were performed near the nucleus and in the periphery to determine the statistical significance of the differences in these regions of interest (arrow heads). In A, C and D, scale bars represent 10 μm. DAPI was used as a nuclear stain (blue). Arrows point to cell protrusions where the proteins co-localize.