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Clear cell renal cell carcinoma (ccRCC) is the most prevalent histological subtype of kidney cancer and is believed to mainly originate from proximal tubular epithelial cells of the nephron.
MTUS1 was significantly downregulated in ccRCC tissues, especially in metastatic tissues. Knockdown of MTUS1 decreased microtubule stability, whereas increased microtubule dynamics by promoting the ratio of unphosphorylated KIF2CS192 adapting to high motility of metastatic cancer cells.
MTUS1 is a potent microtubule-stabilizing protein19, so we assessed the impact of MTUS1 knockdown on microtubule dynamics.
MTUS1 regulates microtubule dynamics via promoting KIF2CS192 phosphorylation by Aurora B.
Western blot and immunohistochemistry further confirmed significantly lower expression of SORBS2 protein in metastatic tissues than that in primary tissues and normal tissues.
Western blot and co-immunoprecipitation (Co-IP):
Wild-type recombinant protein KIF2CWT (1 μg), pointed mutant KIF2CS192A (1 μg), Aurora B (100 ng), and MTUS1 (100 or 500 ng) (recombinant proteins were synthesized and purified by Medicilon) were incubated with 0.2 μCi/μl [γ-32P]-ATP and 30 μM non-radioactive ATP for 30 min at 37°C.
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