Pcdna4c hbrd4

BRD4 siRNA (5′‐CCUGAUUACUAUAAGAUCAdTdT‐3′) was designed and synthesized by Biotend. BRD4‐siRNA or NC‐siRNA was transfected into the cells plated in 6‐well plates at 30% density using Lipofectamine 2000 (Invitrogen) according to the manufacturer's protocol. The pcDNA4c hBrd4 was obtained from Addgene (Plasmid 14441) and the amino acid change T249P was modified by Longqian Biotech. BRD4 plasmids (Longqian Biotech) or control plasmids were transfected into cells by using Viafect Transfection Reagent (Promega).

BRD4 siRNAs were designed and synthesized by Biotend (Shanghai, China). The siRNAs targeting human BRD4 were: BRD4-siRNA1: 5′-CCUGAUUACUAUAAGAUCAdTdT-3′, BRD4-siRNA2: 5′-CAGUGACAGUUCGACUGAUdTdT-3′. Cells were plated onto 6-well plates at 30% density and transfected with BRD4-siRNA or NC-siRNA using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's protocol.
The pcDNA4c hBrd4 was obtained from Addgene (Plasmid 14441), and the amino acid change T249P was modified by Longqian Biotech. Viafect Transfection Reagent was used for transfection. BRD4 expression level was detected by qRT-PCR and western blotting.
pGMLV-SC5 RNAi vectors encoding BRD4 (5′-CCTGATTACTATAAGATCA-3′) were constructed by Genomeditech (Shanghai, China). NOZ cells were infected at a multiplicity of infection (MOI) of 80 in complete culture medium for 24 h. qRT-PCR and western blotting were used to examine the knockdown efficiency.