Figure S4.
LICs act redundantly to recruit dynein and dynactin to RILP-positive late endosomes, and recruitment requires helix 1. (A–D) Cells were depleted of LICs individually or together for 48 h using a total of 20 nM siRNA then transfected with GFP-RILP and fixed 1 d later and labeled for IC (A) or dynactin p150 (B). Control cells (not siRNA treated) were transfected with GFP-RILP. DeltaVision deconvolved images are shown as z-stack projections. Scale bars = 10 μm in the main image, 0.1 µm in the inset, applicable to all images. The percentage of cells with GFP-RILP structures labeled with IC (C) or p50 (D) was scored (± SD). At least 100 cells were scored in each of the three independent experiments. *** = P < 0.001, **** = P < 0.0001, one-way ANOVA with Dunnett’s post-hoc test. (E) HeLaM cells were cotransfected with HA-RILP and GFP-LIC1 full-length, CT2, or CT3. Wide-field images are shown; scale bar = 10 µm.

LICs act redundantly to recruit dynein and dynactin to RILP-positive late endosomes, and recruitment requires helix 1. (A–D) Cells were depleted of LICs individually or together for 48 h using a total of 20 nM siRNA then transfected with GFP-RILP and fixed 1 d later and labeled for IC (A) or dynactin p150 (B). Control cells (not siRNA treated) were transfected with GFP-RILP. DeltaVision deconvolved images are shown as z-stack projections. Scale bars = 10 μm in the main image, 0.1 µm in the inset, applicable to all images. The percentage of cells with GFP-RILP structures labeled with IC (C) or p50 (D) was scored (± SD). At least 100 cells were scored in each of the three independent experiments. *** = P < 0.001, **** = P < 0.0001, one-way ANOVA with Dunnett’s post-hoc test. (E) HeLaM cells were cotransfected with HA-RILP and GFP-LIC1 full-length, CT2, or CT3. Wide-field images are shown; scale bar = 10 µm.

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