Figure 5. RILPL1 depletion reduces JIP4... | Rockefeller University Press

$RILPL1 depletion reduces JIP4 presence on LRRK2-positive lysosomes. (A) U2OS cells were treated with NTC or siRILPL1 RNA. 24 h later, cells were transfected with HaloTag-LRRK2 for 36 h. Then, cells were treated with LLOME (2 h) and were fixed and stained with JIP4 and LAMP2 antibodies. (B) LRRK2+ lysosomes per cell were quantified. Data are mean ± SEM (n = 33–35 cells). Unpaired t test. (C) JIP4+ lysosomes per cell were quantified. Data are mean ± SEM (P = 0.0202, n = 33–35 cells). Unpaired t test. (D) The fraction of LRRK2+/JIP4+ lysosomes compared with LRRK2+/JIP4− lysosomes in NTC vs siRILPL1-treated cells. Data are mean ± SEM (P = 0.0035, n = 31–33 cells). Unpaired t test with Welch’s correction for unequal variance. (E) Cartoon depicting the role of RILPL1 in clustering LRRK2+ lysosomes toward the centrosome and its effect on JIP4 recruitment. Arrowheads indicate LRRK2+/JIP4- lysosomes. Scale bar = 10 µm. NTC, nontargeting control.$

Figure 5.

RILPL1 depletion reduces JIP4 presence on LRRK2-positive lysosomes. (A) U2OS cells were treated with NTC or siRILPL1 RNA. 24 h later, cells were transfected with HaloTag-LRRK2 for 36 h. Then, cells were treated with LLOME (2 h) and were fixed and stained with JIP4 and LAMP2 antibodies. (B) LRRK2⁺ lysosomes per cell were quantified. Data are mean ± SEM (n = 33–35 cells). Unpaired t test. (C) JIP4⁺ lysosomes per cell were quantified. Data are mean ± SEM (P = 0.0202, n = 33–35 cells). Unpaired t test. (D) The fraction of LRRK2⁺/JIP4⁺ lysosomes compared with LRRK2⁺/JIP4⁻ lysosomes in NTC vs siRILPL1-treated cells. Data are mean ± SEM (P = 0.0035, n = 31–33 cells). Unpaired t test with Welch’s correction for unequal variance. (E) Cartoon depicting the role of RILPL1 in clustering LRRK2⁺ lysosomes toward the centrosome and its effect on JIP4 recruitment. Arrowheads indicate LRRK2⁺/JIP4- lysosomes. Scale bar = 10 µm. NTC, nontargeting control.

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