Fig 1: Nox2 (NADPH oxidase-2)-dependent upregulation of GDF6 (growth differentiation factor 6). A, GDF6 levels in conditioned medium of primary cardiac fibroblasts infected with adenoviruses expressing shRNA (short hairpin RNA) targeted against Nox2 (Ad.shNox2) or adenoviruses expressing shRNA targeted against a GFP (green fluorescent protein) control (Ad.shGFP; control) and treated with ANG II (angiotensin II) or vehicle. Representative immunoblot shown at the top and mean data at the bottom; n=3 per group. Ponceau staining of the membrane is shown in Figure XI in the Data Supplement. B, GDF6 protein levels in aorta from Fibro-Nox2KO or matched control (Nox2Flox) mice chronically treated with ANG II or saline vehicle. Representative immunoblot shown at the top and mean data at the bottom; n=4 per group. β-actin was used as a loading control. Densitometry values for GDF6-specific bands were divided by the corresponding densitometry of Ponceau bands in A and β-actin bands in B. Arbitrary units were calculated by normalizing data vs the control group (ie, fold-change). C, GDF6 immunostaining in aorta from Fibro-Nox2KO and control mice treated with ANG II or saline. Representative images are shown on the left and mean fluorescence intensity on the right. GDF6, red; 4′-6-diamidino-2-phenylindole (DAPI), blue; autofluorescence (AF), green. Scale bars=50 µm. 1-way ANOVA followed by Tukey post-test. L indicates vessel lumen. *P<0.05.
Fig 2: GDF6 (growth differentiation factor 6) induces vascular smooth muscle cell (VSMC) growth. A, Concentration-dependent effect of recombinant GDF6 (0–4 μg/mL) on BrdU (bromodeoxyuridine) incorporation in VSMC; n=4–6 per group. B, Representative Western blot for GDF6 expression after silencing using siRNA (silencing RNA) against GDF6 or scrambled control (control) in primary cardiac fibroblasts. C and D, VSMC growth assessed by Ki67 staining after incubation with conditioned media of aortic (C) or cardiac (D) fibroblasts treated with ANG II (angiotensin II). Some fibroblasts were treated with siGDF6 (silencing RNA against GDF6) to reduce GDF6 levels. 1-way ANOVA followed by Tukey post-test (A and C) or Kruskal-Wallis followed by Dunn post-test (D). *P<0.05.
Supplier Page from MyBioSource.com for GDF6 siRNA (Mouse)