Fig 1.
EGFP expression in H9C2 Cells of Group A-E at 48 h post-transfection (200×).
(A): untreated group; (B): treated with pEGFP-N plasmid DNA; (C): treated with PEI/DNA complexes; (D): treated with PEI/DNA complexes with ultrasonic exposure; (E): treated with PEI/DNA complexes plus PFPMS with ultrasonic exposure.
Fig 2.
Effects of AI and MC on cell transfection efficiency and cell viability.
(A1-A2): transfection efficiency and cell viability at different MC (100 μl/ml, 200 μl/ml, 300 μl/ml and 400 μl/ml) when AI was 0.5W/cm2. (B1-B2): transfection efficiency and cell viability at different MC (100 μl/ml, 200 μl/ml, 300 μl/ml and 400 μl/ml) when AI was 1.0 W/cm2. (C1-C2): transfection efficiency and cell viability at different MC(100 μl/ml, 200μl/ml, 300μl/ml and 400μl/ml) when AI was 1.5W/cm2. (D1-D2) transfection efficiency and cell viability at different MC (100 μl/ml, 200 μl/ml, 300 μl/ml and 400 μl/ml) when AI was 2.0 W/cm2. Indication: Symbols indicate significant differences (P < 0.05) from the 100μg/ml. (*), from 200μg/ml (†) and from 300μg/ml (‡)respectively.
Fig 3.
The effect of ET on cell transfection efficiency and cell viability.
(A): transfection efficiency at different exposure time; (B): cell viability at different exposure time. Indication: Symbols indicate significant differences (P< 0.05) from ET of 15 s (*) or 30 s (†).
Fig 4.
The effect of PC on transfection efficiency and cell viability.
A-D showing EGFP expression at PC 5 μg/ml,10 μg/ml,15 μg/ml, 20 μg/ml in sequence. E showing transfection efficiency at different PC with when MC (30%), AI (1.5 W/cm2), and ET(30 s) were fixed. F cell viability at different PC with when MC (30%), AI (1.5 W/cm2), and ET (30 s) were fixed. Indication: Symbols indicate significant differences (P< 0.05) from ET of 5 μg/ml(*) or 10μg/ml (†).
Fig 5.
Effects of shPHD2 delivery on PHD2, HIF-1a, VEGF, bFGF and TGF-β1 mRNA and protein expression in H9C2 cells.
A-E: Expression of PHD2, HIF-1,VEGF, TGF-β1and bFGF was determined by quantitative polymerase chain reaction(qPCR) in H9C2 cells of Normoxic & EGFP, Normoxic & shPHD2-EGFP, Hypoxic & EGFP and Hypoxic & shPHD2-EGFP group presented in the bar graph (asterisk indicates P-value<0.05).
Fig 6.
Effects of shPHD2 delivery on PHD2, HIF-1a,VEGF, bFGF and TGF-β1 protein expression in H9C2 cells.
A1, B1, C1 and D1: Representative Western-blot, showing bands of PHD2, HIF-1α,VEGF, bFGF and TGF-β1 in myocardial specimens collected from the indicated groups (asterisk indicates P—value<0.05). A2-A3,B2-B3,C2-C4 and D2-D4:Densitometric measurements of the PHD2, HIF-1α, VEGF, bFGF and TGF-β1 to GAPDH and presented in the bar graph (asterisk indicates P-value<0.05). Indication: Normoxic & EGFP: normoxic H9C2 cells transfected with EGFP plasmid; Normoxic & shPHD2-EGFP: normoxic H9C2 cells transfected with shPHD2-EGFP plasmid; Hypoxic & EGFP: hypoxic H9C2 cells transfected with EGFP plasmid; Hypoxic & shPHD2-EGFP: hypoxic H9C2 cells transfected with shPHD2-EGFP plasmid.