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. 2003 Nov 1;112(9):1395–1406. doi: 10.1172/JCI17700

Figure 8.

Figure 8

(a–c) Neonatal rat cardiac myocytes were treated with either control Morpholino oligo or Morpholino antisense oligo for rat Trx1 (2 μM) using osmotic delivery. Myocytes were harvested 48 hours after application of the oligo. (a) Expression of Trx1 was determined by immunoblotting with anti-Trx1 Ab (4H9). (b) Total cardiac myocyte protein content was determined. The mean value in control Morpholino oligo–treated myocytes was designated as 1. (c) Cellular content of MDA and MDA plus 4-HAE was determined as described in Methods. (d) COS-7 cells grown in 60-mm dishes were transfected with either empty vector plasmid or pcDNA3.1 harboring DN-hTrx1, antisense rat Trx1 (AS-Trx1), or Trx1. Forty-eight hours after transfection, cells were harvested, and expression of Trx1 was determined by immunoblot analysis. The level of Trx1 expression quantitated by densitometric analyses is shown. (e) Cardiac myocytes were transfected with ANF-luciferase (-638) (ANF-luciferase reporter gene containing 638 base pairs upstream of the rat ANF gene transcription start site) and SV40-β-galactosidase, together with pcDNA3.1 alone (empty vector) or with pcDNA3.1 harboring DN-hTrx1, antisense rat Trx1, or Trx1. Forty-eight hours after transfection, activities of luciferase and β-galactosidase were determined. The activity of luciferase was normalized by that of β-galactosidase.