Fig. 3

GAB1 is a target gene of miR-29a. A, Predicted binding site between miR-29a and GAB1. B, The binding of miR-29a to GAB1 verified by dual-luciferase reporter gene assay. ^* p < 0.05 vs. the cells co-transfected with mimic-NC and pMIR-GAB1-Wt plasmids or pMIR-GAB1-Mut plasmids (the mimic-NC group). C&D, Western blot analysis of GAB1 protein in MLE-12 cells of each group. ^* p < 0.05 vs. MLE-12 cells transfected with mimic-NC (the mimic-NC group). ^# p < 0.05 vs. MLE-12 cells delivered with inhibitor-NC (the inhibitor-NC group). E&F, the positive expression of GAB1 protein in lung tissues of each group detected by immunohistochemistry (× 400). ^* p < 0.05 vs. neonatal mice under exposure of RA (the RA group). ^# p < 0.05 vs. neonatal mice with hyperoxia-induced BPD receiving miR-29a antagomir administration (the BPD + miR-29a antagomir group). N = 15. Measurement data were expressed as mean ± standard error. Data comparison between two groups was examined by unpaired t-test. The experiment was repeated 3 times independently. miR-29a, microRNA-29a; GAB1, GRB2-associated-binding protein 1; BPD, bronchopulmonary dysplasia; RA, room air; Wt, wild type; Mut, mutant; NC, negative control