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The study built on many years of research that has illuminated the versatility of USP8.
"We had previously reported that USP8 regulates pituitary hormone secretion," said Fukushima, referring to a paper published in Nature Genetics in 2015.
It was this 'accidental' finding, combined with promising results from other groups in the US that led the team to examine the role of USP8 in the formation of COPII carriers.
In research tracing back more than a decade, Komada and others have clarified the conventional role of USP8 in the regulation of endocytosis5.
After that we changed the medium to cell culture medium with 2% FBS with indicated concentrations of USP8 inhibitor for 24 and 48 h.
The medium with indicated concentrations of USP8 inhibitor was replaced every 3 or 4 days.
The cells were incubated for 4 and 24 h with the indicated concentrations of USP8 inhibitor.
After exposure to USP8 inhibitor for 24 and 48 h, cells were detached and then washed once with cold PBS, suspended in 1 x binding buffer at a concentration of 5 x 10[sup]5 cells/ml.
To investigate that targeting USP8 with its specific inhibitor might exhibit an anticancer effect in the corticotroph adenomas, we first examined the effect of USP8 inhibitor on downstream protein levels including EGFR, ERBB2, and Met.
To determine the specificity of USP8 inhibitor effects, cell viability was assessed in Hepa 1-6, HEK293T, and PC12 cell lines after 24 h treatment without or with increasing concentration of USP8 inhibitor (1–10 [micro]mol/L).
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