Wnt5A decreases ROR1 expression through proteasomal degradation.

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Wnt5A decreases ROR1 expression through proteasomal degradation. Wnt5A decreases ROR1 expression through proteasomal degradation. A, RT-PCR analysis of ROR1 mRNA levels in Wnt5A-low UACC1273 and G361 cell lines ± treatment with rWnt5A (200 ng/mL, 16 hours; ***, P < 0.001; error bars, SD). B, surface biotinylation and Western blot analyses of ROR1 protein expression in UACC1273 cells (± rWnt5A) as well as metastatic UACC903, M93-047, and UACC647 cell lines. C, time course analysis (5 minutes, 1 hour, and 16 hours) of ROR1, Wnt5A, and HSP90 in G361 and UACC1273 cells. D, expression of ROR1 (green) protein in G361 cells following pretreatment with the proteasome inhibitor (MG132, 10 μmol/L, 1 hour) in the presence or absence of rWnt5A (200 ng/mL, 10 minutes). E, Western blot analysis of ROR1 protein in UACC1273 cells following pretreatment with the proteasome inhibitor (MG132, 10 μmol/L, 1 hour) in the presence or absence of rWnt5A (200 ng/mL, 16 hours). F, Western blot analysis of ROR1 in poorly invasive UACC1273 cells following treatment with the PKC inhibitor GO6983 (1 μmol/L, 17 hours). G, Western blot analysis of ROR1 protein in UACC1273 cells following pretreatment with the PKC inhibitor GO6983 (1 μmol/L, 17 hours) in the presence or absence of rWnt5A (200 ng/mL, 16 hours). DAPI, 4′,6-diamidino-2-phenylindole. Michael P. O'Connell et al. Cancer Discovery 2013;3:1378-1393 ©2013 by American Association for Cancer Research