Figure 3.

TRAF3 is the target gene of miR‐29b‐3p. (a) The expression levels of miR‐29b‐3p after H/R injury in IEC‐6 cells and II/R injury in mice based on real‐time PCR assay. (b) The diagram of miR‐29b‐3p conservative seed binding sites on the 3′‐UTRs of the target genes (TRAF3). (c) Relative luciferase expression with TRAF3 3′‐UTR after co‐transfected with miR‐29b‐3p mimic or NC. IEC‐6 cells were seeded in 24‐well plates and co‐transfected with miR‐29b‐3p mimic or NC and TRAF3‐WT or TRAF3‐mutant (MUT). After 24 hr of transfection, the luciferase activities were assessed using Dual‐Luciferase Reporter Assay Kit. (d, e) The expression levels of TRAF3 after co‐transfection. IEC‐6 cells were seeded in six‐well plates for 24 hr and then co‐transfected with miR‐29b‐3p mimic NC and TRAF3 overexpression plasmid NC, miR‐29b‐3p mimic and TRAF3 overexpression plasmid, single transfected with TRAF3 overexpression plasmid. Then the mRNA level of TRAF3 was assessed by real‐time PCR assay, and the protein level of TRAF3 was detected by western blotting assay after transfection. All data are expressed as the mean ± SD (n = 5). *P < .05 compared with control group in Figure 3a,d,e and compared with NC group in Figure 3c; ^# P < .05 compared with 120 min of reoxygenation or 90 min of reperfusion group in Figure 3a and compared with TRAF3 plasmid group in Figure 3d,e; n.s., no significance