Transduction efficiency after intramyocardial injection of NGF siRNA <i>in vivo</i>.

<p>(A) Representative site of intramyocardial injection (arrowed) and GFP expression in infarcted hearts of rats in the MI-SiNGF group at 1 wk time point. (B) Relative expressions of NGF mRNA, detected by real-time quantitative RT-PCR, in the sham-operated, MI-control, MI-GFP and MI-SiNGF groups at various time points (1, 2, 4 and 8 wk) after intramyocardial injection of NGF siRNA. Relative gene expressions of NGF were analyzed by the 2^-ΔΔCT method taking those in the sham-operated groups as 1. (C) Expressions of NGF (27 KDa) and GAPDH (36 KDa), analyzed by Western blot, in the sham-operated, MI-control, MI-GFP and MI-SiNGF groups at various time points (1, 2, 4 and 8 wk) after intramyocardial injection of NGF siRNA. (D) Relative protein expressions of NGF. The protein expression levels in (C) were quantified with Quantity AlphaEaseFCTM imaging software. Relative expression of NGF was normalized to GAPDH. Data were presented as mean ± SD. *<i>p<0.05</i> MI-control group vs. sham-operated groups, †<i>p<0.05</i> MI-SiNGF group vs. MI-control or MI-GFP groups. The results showed that expression levels of NGF mRNA and protein were induced in the MI-control group compared to the sham-operated group. NGF mRNA and protein in the MI-GFP group had no significant levels compared to the MI-control group, while those in the MI-SiNGF group were reduced compared to the MI-GFP group.</p

Histological and RT-PCR results of TH- and GAP 43 expression.

<p>Data were expressed as means ± SD.</p><p>* <i>P<0.05</i> vs. Sham-operated group;</p>†<p><i>P<0.05</i> vs. MI-GFP group.</p

Representative images for infarcted hearts at 8 wk time point after MI.

<p>(A) Macroscopical view of the infarcted heart. (B) Masson’s trichrome staining of infarcted area.</p

Echocardiographic and Hemodynamic data at 8 wk after transduction.

<p>Values were means±SD. LVIDd, left ventricular internal diameter at end-diastolic phase; LVIDs, left ventricular internal diameter at end-systolic phase; EF: left ventricular ejection fraction; FS: left ventricular fractional shortening; HR, heart rate; MAP: mean arterial pressure; LVEDP: LV end-diastolic pressure; LVESP: LV end-systolic pressure; dP/dt_max: the maximal rates of increase in left ventricular pressure; dP/dt_min: the maximal rates of decrease in left ventricular pressure.</p><p>* <i>P<0.05</i> vs. Sham-operated group;</p>†<p><i>P<0.05</i> vs. MI-GFP group.</p

Additional file 1 of Lung-specific RNA interference of coupling factor 6, a novel peptide, attenuates pulmonary arterial hypertension in rats

Figure S1. Validation of the MCT-induced PAH model. Figure S2. Validation of the MCT plus - pneumonectomy induced PAH model. Figure S3. CF6 upregulation in human lung cancer tissue. Figure S4. Kaplan-Meier survival curves. (DOCX 4583 kb

Additional file 1: Figure S1. of Role of P2X7R in the development and progression of pulmonary hypertension

Validation of the MCT plus - pneumonectomy induced PAH model. (A-B) Rats were given a single intraperitoneal injection of 60 mg/kg MCT one week after left pneumonectomy or vehicle, and RV systolic pressure (A) and RV weight (B) were measured 1, 2, 3, or 4 weeks after MCT challenge. (C–F) H&E staining and a-SMA staining of lung tissue sections at 4 weeks after MCT injection. (G) The % medial wall thickness was calculated as [(medial thickness × 2)/external diameter] × 100. Scale bars  = 50 μm. All data are expressed as mean ± SD. n =  6 per group. **p < 0.01 and *p < 0.05. Figure S2 Representative double-immunostained images of P2X7R (Red), co-stained for FITC-CD68, DAPI (Blue) for nuclei and merged images in the P/MCT (A), sham group (B) and negative control (C), Original magnification × 40. Scale bar = 50 μm for all images. (DOCX 1872 kb