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We next determined whether transcription is directly regulated by p73. As several p73 antibodies failed to precipitate endogenous p73, HA-tagged p73β was first transfected into cells to facilitate its detection. After gefitinib treatment, the recruitment of p73 to the promoter containing two p53-binding sites was found to significantly increase in a time-dependent manner in JHU-012 and JHU-029 cells. In contrast, the binding of p53 to the same region was unaffected by gefitinib treatment (). Using a series of deletion reporter constructs (), we found that only the reporters containing the two p53-binding sites, such as Frag A, abc and Frag c, were significantly activated by gefitinib treatment (). Furthermore, knockdown of p73 by small interference RNA (siRNA) impaired gefitinib-induced PUMA expression (). These data suggest that p73 activates transcription after gefitinib treatment through the p53-binding sites.
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The season of celebration is under way. Staff recognition events are being held this month for employees retiring, reaching 20-plus years of service and celebrating five-, 10- and 15-year anniversaries. Various members of JHU’s leadership, including President Ron Daniels and Provost Lloyd Minor, have shown their appreciation for the collective years of service by attending […]