Suppression of renal cell carcinoma growth by inhibition of Notch signaling in vitro and in vivo
J. Clin. Invest. Jonas Sjölund, et al. 118:217 doi:10.1172/JCI32086 [Go to this article.]

Figure 1
Notch signaling pathway components are expressed in CCRCC cells and maintained in a HIF-1α– and HIF-2α–independent manner. (A) Immunoblots of SKRC-7, SKRC-10, SKRC-21, SKRC-17, SKRC-52, and Caki-2 cell lysates analyzed for indicated proteins. Actin was used as a control for equal loading of samples. (B) Q-PCR analyses of Jagged-1, Jagged-2, Notch-1, Notch-2, Hes-1, and Hey-1 mRNA expression in CCRCC cells. mRNA levels were normalized to succinate dehydrogenase complex subunit A (SDHA), tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein (YWHAZ), and ubiquitin C (UBC) expression. Data shown are mean + SD of representative experiment performed in triplicate. (C) 786-O, PRC3, and WT7 cell lysates were analyzed for pVHL, HIF-2α, Jagged-1, Notch-1, and Hes-1 expression by Western blot analyses. (D) VEGF, Hes-1, and Hey-1 mRNA levels in WT7, PRC3, and 786-O cells. Data shown are mean + SD of representative experiment performed in triplicate. (E) PRC3 and WT7 cells, transfected with control (c-si) or HIF-2α siRNA (siHIF-2α) for 6 hours and then incubated under either normoxic (21% O₂) or hypoxic (1% O₂) conditions for 24 hours prior to protein extract preparation, were subjected to immunoblotting of indicated proteins. (F) Q-PCR analyses of indicated mRNAs following control or HIF-1α siRNA (siHIF-1α) transfection of SKRC-10 cells. Treatment procedure (N, normoxia; H, hypoxia) and transfection were performed as indicated in E. Data shown are mean + SD of representative experiment performed in triplicate.