Fig. 3: Functionality of transgenic AAT. | Gene Therapy

Fig. 3: Functionality of transgenic AAT.

From: Pulmonary transplantation of alpha-1 antitrypsin (AAT)-transgenic macrophages provides a source of functional human AAT in vivo

Fig. 3

A Formation of a complex between AAT and elastase. AAT present in the MΦ supernatant binds elastase and forms a complex represented as a band with higher molecular weight detected with anti-human AAT antibody in western blot analysis. In addition to the complex also a cleaved form of AAT of approx. 45–50 kD is visible. Same samples were used as for western blot in Fig. 1H. B Elastase inhibition by supernatant of Cbx-EF1α-AAT MΦ. AAT protein at different concentrations was used as a positive control. Supernatant of mock-transduced MΦ served as base for AAT protein and conditioned medium dilutions. n = 3 technical replicates. Lines represent mean ± SD. Statistical analysis was performed using one-way ANOVA with Tukey’s post-hoc test. C Percentage of propidium iodide (PI) positive mAM cells after apoptosis induction with 25 µM Staurosporine (STS). AAT protein at concentrations of 10–200 µg/ml in supernatant (supn.) of mock-transduced MΦ served as positive control. Supernatants of CAG-AAT and Cbx-EF1α-AAT MΦ were added pure, undiluted (1) or in 1:2, 1:4, and 1:8 dilution. Untreated, +STS, AAT 10–200 μg/ml n = 4; CAG-AAT and Cbx-EF1α-AAT 1:8–1 n = 2 biological replicates from independent transductions and 1 technical replicate. Lines represent mean ± SD.

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