Research Paper Volume 12, Issue 20 pp 20198—20211

Silencing of long non-coding RNA MEG3 alleviates lipopolysaccharide-induced acute lung injury by acting as a molecular sponge of microRNA-7b to modulate NLRP3

Figure 3. LncRNA MEG3 binds to miR-7b and competes with NLRP3. RAID and LncBase predicted lncRNAs that might bind to miR-7b (A). RNA-FISH for lncRNA MEG3 expression in NR8383 cells (× 200) (B). Dual luciferase reporter gene assay verified the targeting relationship between miR-7b and NLRP3 (C). RT-qPCR for targeting relation of miR-7b and lncRNA MEG3, * p < 0.05 vs. NC mimic (C). Correlation analysis of miR-7b and lncRNA MEG3 expression (D). The expression of miR-7b after lncRNA MEG3 overexpression by RT-qPCR, * p < 0.05 vs. oe-NC (E). RIP assay for the binding of miR-7b with lncRNA MEG3 or NLRP3 respectively, * p < 0.05 vs. IgG (F). The expression of lncRNA MEG3 in LPS-induced cells was detected by RT-qPCR, * p < 0.05 vs. PBS + NR8383 (G). The NLRP3 expression normalized to GAPDH detected by Western blot analysis, * p < 0.05 vs. si-NC + NC mimic, # p < 0.05 vs. si-MEG3 + NC mimic, & p < 0.05 vs. si-MEG3 + miR-7b mimic (H). Measurement data were expressed as mean ± standard deviation. Comparison between two groups was conducted using unpaired t-test. Data among multiple groups were tested using ANOVA, followed by Tukey’s post hoc test. The experiments were repeated three times independently.