Fig. 7
PDGF-BB-H19-Let-7b axis promotes the proliferation of PASMCs. a: PASMCs were stimulated with IL-1β and PDGF-BB at a concentration of 100 ng/ml for 48 h. b: PASMCs were transfected with 50 nM miCon, let-7b-mimic, iCon or iLet-7b for 24 h and 48 h. c: PASMCs were cotransfected with 1000 ng of vector or pH 19 with 50 nM siCon or siAT1R for 6 h, 12 h, 24 h, 48 h and 72 h, respectively. d: PASMCs were transfected with vector or pH 19 at 0 ng, 50 ng, 100 ng, 500 ng and 1000 ng for 48 h. In a, b, c and d, PASMCs were washed with sterile PBS after transfection or stimulation and cultured in CCK8 diluent for 2 h. Absorbance was detected at 450 mm. e: PASMCs were transfected with 1000 ng of vector or pH 19 for 48 h. f: PASMCs were stimulated with PDGF-BB at a concentration of 100 ng/ml for 48 h. Then, PASMCs were transfected with 50 nM siCon or siH19 for 24 h. H19, Ki67 and PCNA mRNA were detected by RT-qPCR. g: PASMCs were transfected with 1000 ng of vector or pH 19 for 48 h. h: PASMCs were stimulated with PDGF-BB at a concentration of 100 ng/ml for 48 h. Then, PASMCs were transfected with (50 nM siCon + 150 nM iCon), (50 nM siH19 + 150 nM iCon) or (50 nM siH19 + 150 nM iLet-7b) for 24 h. Cell-cycle distribution was analysed by flow cytometry. i: Scratch wound healing assay of PASMCs transfected with 1000 ng of vector or pH 19. Values were presented as means ± SD from 3 independent experiments. *0.01 ≤ P ≤ 0.05 (different from the corresponding control); **0.001 ≤ P ≤ 0.009 (different from the corresponding control); ***P < 0.001 (different from the corresponding control); #0.01 ≤ P ≤ 0.05 [different from the (pH 19 + siCon) group]; ##0.001 ≤ P ≤ 0.009 [(siH19 + iLet-7b) vs. (siH19 + iCon) or (pH 19 + siCon) vs. (pH 19 + siAT1R)]