Nrf2 siRNA (m)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Nrf2 siRNA: 5′ GAAUGGUCCUAAAACACCAtt-3′ siRNA concentration-50 nm Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio). Incubate cells for 24 h at 37°C.

Protocol tips
Nrf2 siRNA: 5′ GAAUGGUCCUAAAACACCAtt-3′ siRNA concentration-50 nm Add diluted siRNA to diluted Lipofectamine® RNAiMAX Reagent (1:1 ratio). Incubate cells for 24 h at 37°C.

Publication protocol

Silencer predesigned small interference RNA (siRNA) for human p62 (siRNA ID s16960), Nrf2 (siRNA ID s9491), and control siRNA (AM4611) were obtained from Ambion (Austin, TX) and used to inhibit p62 and Nrf2 protein. The coding strand of p62 siRNA was 5′-GGAGCACGGAGGGAAAAGAtt-3′; the coding strand of Nrf2 siRNA was 5′-GAAUGGUCCUAAAACACCAtt-3′. Normal BEAS-2B cells and CdT cells were seeded in 96- or 6-well culture plates and transfected with 50 nm siRNA duplexes using LipofectamineTM RNAi MAX (Invitrogen) according to the manufacturer's instructions. Twenty-four hours after transfection, the cells were harvested, and cellular levels of proteins specific for the siRNA transfection were checked by immunoblotting.

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