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Effects of Genistein and Apocynin on Protein Activity: Insights from Akt and Erk1/2 Phosphorylation

This study investigates the impact of Genistein and Apocynin on protein activity, particularly focusing on Akt and Erk1/2 phosphorylation. The findings reveal a 3-fold increase in Akt phosphorylation and a notable modulation in Erk1/2 phosphorylation in response to the treatments. The results shed light on the signaling pathways affected by these compounds and provide valuable insights into their mechanisms of action. Various techniques such as A.B Raf-1 pull-down, Ascorbate supplementation, and IB p21ras labeling were employed to study the time-dependent effects of these compounds on protein activity. The use of NEM, DTT, BHPDP labeling, and SA pull-down further enhanced the understanding of the molecular changes induced by Genistein and Apocynin. Overall, this research demonstrates the potential therapeutic implications of these compounds in modulating protein function.

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Effects of Genistein and Apocynin on Protein Activity: Insights from Akt and Erk1/2 Phosphorylation

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  1. AG1478 - - - - - - + + Genistein - - - - + + - - P. Toxin - - + + - - - - ONOO- - + - + - + - + 62 kDa-- Akt-P 4 42/44 kDa- Erk1/2-P ** C 3 Fold Increase Apocynin - - - - + + 2 L-NAME - - + + - - ONOO- - + - + - + 1 62 kDa--- Akt-P 0 Control BGSH BGSH SA 42/44 kDa- Erk1/2-P Supplemental Figures A B

  2. Raf-1 Pull-down Ascorbate - - - + Time: 0 5 15 0 5 15 min DTT - - + - ONOO- - + + + IB p21ras ---21 kDa IB p21ras --21 kDa Total Lysate Desalted proteins NEM, DTT, BHPDP labeling, SA pull-down D ONOO- - 100 µM 1mM IB p21ras ---21 kDa E F

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