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. 2018 Jun 22;293(33):12719–12729. doi: 10.1074/jbc.RA118.003075

Figure 3.

Figure 3.

KIF5B mediates AR membrane transport. A and B, the effects of KIFC3-tail and KIF5B-tail on AR membrane translocation. LNCaP cells were transiently transfected with KIFC3-tail or KIF5B-tail for 48 h, and treated with 1 nm R1881 for 20 min. TS/TI fractions were extracted and analyzed by Western blotting. Membrane AR levels were calculated as AR/Giα3 ratios and expressed relative to that of the leftmost group. C, KIF5B-tail, but not KIFC3-tail, interferes with rapid activation of AKT. LNCaP cells were transfected with KIF5B-tail or KIFC3-tail, and treated with 1 nm R1881 treatment for 30 min. Whole cell lysates were collected and probed with antibodies against p-AKT and AKT. D, KIF5B knockdown. After LNCaP cells were transfected with siRNAs for 48 h, lysates were analyzed by Western blotting to confirm the knockdown of KIF5B. E, KIF5B knockdown interrupts AR membrane translocation. LNCaP cells were transfected siRNAs for 48 h and treated with 1 nm R1881 for 20 min. TS/TI fractions were extracted and analyzed for AR distribution. Left, representative blots are shown. Right, AR levels in TI fractions were calculated as the AR/Giα3 ratios and expressed relative to that of the leftmost group. The mean ± S.D. from three experiments are plotted. *, p < 0.05.