Stem Cells International / 2014 / Article / Fig 1

Research Article

Reducing TRPC1 Expression through Liposome-Mediated siRNA Delivery Markedly Attenuates Hypoxia-Induced Pulmonary Arterial Hypertension in a Murine Model

Figure 1

In vitro and in vivo pilot studies validating the murine model of pulmonary arterial hypertension (PAH). (a) Immunofluorescent studies demonstrating successful primary cell culture using the aortic smooth muscle cells (SMCs) from murine aorta, showing SMCs (white arrows) and non-SMCs. (b) Western blot analysis () of primary SMC culture showing markedly suppressed transient receptor potential cation channel (TRPC) 1 protein expression in SMCs after siRNA-knockdown (20 nM), suggesting successful knockdown of TRPC1 expression with Lipofectamine delivered siRNA. (c) Significantly elevated right ventricular systolic pressure (RVSP) in animals by 14 days after hypoxia treatment compared to that in normal controls, indicating successful establishment of the murine model of pulmonary arterial hypertension. versus control, ( for each group). (d) Western blot and (e) IF analyses () of lung tissues demonstrating successful pulmonary delivery of Lipofectamine delivered enhanced green fluorescence protein plasmid (pEGFP) through inhalation. (d) Western blot analysis showing enhanced EGFP expression as pSuper-EGFP concentration increased. (e) IF demonstrating (B) control plasmid without EGFP expression in lung tissue and (H) the merged image, indicating successful Lipofectamine transfection of EGFP plasmid into the pulmonary arteriolar (PA) SMCs (white arrows). Blue fluorescence indicates DAPI-stained nuclei. α-SMA: alpha-smooth muscle actin staining.

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