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. 2014 Jun 4;9(6):e99056. doi: 10.1371/journal.pone.0099056

Figure 7. CXCR6 was effectively down-regulated by siRNA techonology.

Figure 7

To silence the CXCR6 gene expression, we transfected A549 cells with phU6/GFP/Neo plasmid containing short hairpin RNA (shRNA) molecules targeted against CXCR6, with the usage of Lipofectamine 2000 (Invitrogen). The sequences for three shRNA oligonucleotides were: (CXCR6-2819-1): 5′-ctGAG GAC AAT TCC AAG ACT T-3′ (sense) and 5′-AAG TCT TGG AAT TGT CCT CAG-3′ (Anti-sense); (CXCR6-2820-2) 5′-ctCAC CAT GAT TGT CTG CTA T-3′ (sense) and 5′-ATA GCA GAC AAT CAT GGT GAG-3′ (Anti-sense); (CXCR6-2821-1) 5′-gcTTG CTC ATC TGG GTG ATA T-3′ (sense) and 5′-ATA TCA CCC AGA TGA GCA AGC-3′ (Anti-sense). Efficiency of RNA interference against CXCR6 was validated by western blot. It was shown in Fig. 7 that CXCR6 protein was substantially expressed in A549 cells (bank-control, without any treatment) and the RNA interference technology effectively inhibited CXCR6 expression in A549 cells. Lane 1: Blank-control; Lane 2: RNA-control; Lane 3: CXCR6 shRNA-(2821-1); Lane 4: CXCR6 shRNA-(2820-2); Lane 5: CXCR6 shRNA-(2819-1).