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. 2014 Dec 19;46(12):e126. doi: 10.1038/emm.2014.86

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Copyright © 2014 KSBMB.

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PA does not affect HPH activity. (a) Total RNA in vector (Vec)- and PLD-transfected cells (PLD) was isolated by the TRIzol reagent and was subjected to Q-PCR for analysis of HIF-1α mRNA. (b) A VHL capture assay using biotinylated HIF peptide was performed as described under Materials and methods. The assay was performed in the presence of the indicated concentrations of PA and resultant blots were probed for FLAG (VHL). Phenanthroline (100 μM), an iron chelator, was used as a positive control. (c) Renal carcinoma cells that are deficient for VHL function (UMRC2) or a clonally selected line with stably expressing VHL (UMRC2/VHL) were treated with PA for 1 h and HIF-1α protein was detected in nuclear extracts.