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We generated a Nox4D mutant with a single amino acid substitution in the NADPH-binding domain (proline 130 to histidine, Figure 10A) that would be expected to impair its activity based on previous studies with Nox4 and Nox2. gosloto login 27,28 VSMCs were transfected with wild-type or mutant Nox4D (Nox4DM) and protein expression confirmed by immunoblotting