Fig 7. The effects of lncRNA-GAS5 mediated by Annexin A2.

A2 = Annexin A2; Mock = control group, only empty plasmid transfected; NC = Negative control group, only control siRNA transfected; A2 siRNA = Annexin A2 knockdown group transfected with silence siRNA; A2 over = Annexin A2 overexpression group transfected with Annexin A2 expression plasmid; GAS5 down = lncRNA-GAS5 knockdown group transfected with silence siRNA. A:Silver stains for protein gels obtained by lncRNA-GAS5 RNA Pulldown. Bio-GAS5 (sense): treatment group; Bio-GAS5 (antisense): control groups; GAS5 (sense): negative control groups. The GAS5- specific-binding protein gels (in Red box) was identified by MALDI-TOF-MS. Results show that Annexin A2 isoform 2 is a direct binding protein to lncRNA-GAS5. N = 2. B: The Annexin A2 expression level was knockdowned in HSVSMCs effectively by siRNA. Values are mean±SE, N = 3; *, P<0.05. C: The Annexin A2 expression level was over-expressed by constructing and transfecting expression plasmid. Values are mean±SE, N = 3; *, P<0.05. D: The proliferation abilities of HSVSMCs were reflected indirectly by the OD450 values measured using CCK-8 kit when Annexin A2 expression level were knockdowned or over-expressed in HSVSMCs. Values are mean±SE, N = 3; *, P<0.05. E: The proliferation abilities of HSVSMCs were reflected indirectly by the OD450 values measured using CCK-8 kit when Annexin A2 and lncRNA-GAS5 expression levels were knockdowned or over-expressed simultaneously in HSVSMCs. Firstly Annexin A2 siRNA or expression plasmid were transfected into the HSVSMCs, six hours later, the cell culture medium were changed and then lncRNA-GAS5 siRNA were transfected into the HSVSMCs. Another 48 hours later, the OD450 values were measured by CCK-8 kit. Values are mean±SE, N = 3; *, P<0.05.