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. 2017 Feb 14;7:42498. doi: 10.1038/srep42498

Figure 6. Lnc-MKI67IP-3 inhibited the pro-inflammatory effects of let-7e as a ceRNA.

Figure 6

HUVECs were respectively treated as shown in the figure for 24 hours. Subsequently, various assays were performed. (a) The mRNA expression of IκBβ was assayed by qPCR. HUVECs transfected with empty vector were used as controls. *P < 0.05 vs. all other groups except for the MKI67IP-3-wt + mimic NC group; △P < 0.05 vs. all other groups except for the let-7e + MKI67IP-3-mut (lacking the binding site of let-7e) group; †P < 0.05 vs. all other groups except for let-7e group; ★P < 0.05 vs. all other groups except for MKI67IP-3-wt group. (b) IκBβ protein in the cytoplasm and NF-κB p65 protein in the cytoplasm and nucleus was detected using Western blotting. The results showed that lnc-MKI67IP-3 could antagonize the effects of let-7e on the IκBβ expression and nuclear translocation of NF-κB p65. Full-length blots are presented in the Supplementary Figure S5. (c) In the culture medium of each group, the secretion of five important inflammatory cytokines and adhesion molecules, transcriptionally regulated through NF-κB p65, was detected using ELISA. HUVECs transfected with empty vector were used as controls. *p < 0.05 vs. all other groups except for let-7e+ MKI67IP-3-Mut group. △p < 0.05 vs. all other groups except for let-7e group. (d) Lnc-MKI67IP-3 expression in HUVECs with different treatments was detected using qPCR. The untreated HUVECs were used as controls. *P < 0.05 vs. all other groups. (e) The expression of mature let-7e and lnc-MKI67IP-3 in the cytoplasm (cyto) and whole HUVECs (whole) were detected using qPCR.