Fig. 2: DENND6A promotes juxtanuclear clustering of lysosomes.

a Schematic representation of the quantification of cumulative intensity distribution method for lysosomal distribution (adapted from⁴²). b Unstarved HeLa cells expressing either GFP alone or DENND6A-GFP were fixed, stained with LAMP1 antibody and imaged using confocal microscopy (Leica SP8). The cell periphery is outlined by a white dotted line. Scale bar = 10 µm for low magnification images; 3.4 µm and 2.1 µm for high magnification images corresponding to inset 1 and 2. c Quantification of cumulative distribution of LAMP1 intensity; mean ± SEM; two-tailed extra sum of F-squares test following nonlinear regression and curve fitting; n = 30 cells from 3 replicates. d Immunoblot showing the protein levels in WT and DENND6A KO HeLa cells. Immunoblot probed with anti-DENND6A and anti-GAPDH antibodies. e WT and DENND6A KO unstarved HeLa cells were fixed, stained with LAMP1 antibody and DAPI, and imaged using confocal microscopy (Leica SP8). The cell periphery is outlined by a white dotted line. Scale bar = 10 µm. Yellow arrow indicates peripheral lysosomes.