FIGURE 2:

Knockdown and rescue of sortilin-dependent, Mn-induced GPP130 redistribution to lysosomes. (A) Control and sortilin knockdown cells were subjected to a 0, 1, or 2 h Mn treatment (500 μM) and fixed and stained using anti-GPP130 (red) and anti-giantin (green) antibodies. Merged and thresholded (GPP130 only) images are also shown for visualization of GPP130 punctae. Bar = 5 µm. (B) Quantification shows the number of GPP130 punctae at each time point of Mn treatment (n = 3 ± SEM, >15 cells per experiment, *p < 0.0006). (C) To assess rescue, control and sortilin knockdown cells were transfected with a siRNA immune version of the sortilin-myc plasmid or an empty vector. The cells were treated 24 h post–plasmid transfection with 2 h of Mn treatment and then were stained using anti-GPP130 (red) and anti-myc (green) antibodies. Bar = 5 µm. (D) The average number of punctae after the 2 h Mn treatment is quantified for control and knockdown cells (± SEM, n = 3, >10 cells per experiment, *p < 0.002).