IκB-Kinase-epsilon (IKKε) over-expression promotes the growth of prostate cancer through the C/EBP-β dependent activation of IL-6 gene expression


Supplementary Materials


Supplementary

Primers were described as follows: 5ʹ-3ʹ, mutations were written in lower case and underlined. 



  




  




  


Table S1 : Primers used for IKKε gene cloning, mutation of IL-6 promoter transcription binding sites, generation of the pORF9-C/EBP-β-Flag 2 plasmid and ChIP assays Primer sequences

 S1 
	IKKε	Forward Reverse	CACCGGTCAGATGCAGAGCACA GCTCAGACATCAGGAGGT
	CREB-1	Forward Reverse	CTTGCCATGCTAAAGtgtGTCACATTGCACAATCTTAATAAGG CCTTATTAAGATTGTGCAATGTGACacaCTTTAGCATGGCAAG
	CREB-2	Forward Reverse	CTTGCCATGCTAAAGGACGTtggATTGCACAATCTTAATAAGG CCTTATTAAGATTGTGCAATccaACGTCCTTTAGCATGGCAAG
	CEBP-1	Forward Reverse	GCTAAAGGACGTCgagTTGCACAATCTTAATAAGGTTTCC GGAAACCTTATTAAGATTGTGCAActcGACGTCCTTTAGC
	CEBP-2	Forward Reverse	GCTAAAGGACGTCACATTttcCAATCTTAATAAGGTTTCC GGAAACCTTATTAAGATTGgaaAATGTGACGTCCTTTAGC
	CEBP-3	Forward Reverse	GCTAAAGGACGTCACATTGCACAggaTAATAAGGTTTCC GGAAACCTTATTAAtccTGTGCAATGTGACGTCCTTTAGC
	Flag 2 -SphI	Forward Reverse	GATTACAAGGATGACGACGATAAGCACATG TGCTTATCGTCGTCATCCTTGTAATCCATG
	IL-6 ChIP	Forward Reverse	GTGTCTTGCCATGCTAAAG AGACATCTCCAGTCCTATATT

Supplementary Figure S1: The IKKε effect on PC3 cell proliferation after addition of exogenous IL-6. Exogenous IL-6 (50 ng/mL) was added to the low-serum culture medium of PC3-6TR-shIKKε cells. Cell growth was measured using the real-time IncuCyte TM cell proliferation assay (Essen BioScience Inc.). Each measurement was performed in triplicate and each experiment was repeated twice. Error bars represent the standard error of the mean. 

Supplementary