Figure 6. Silencing HO-1 and NQO1 induction in human monocytes enhances LPS-induced inflammatory responses.
THP-1 cells were transfected with 30 nM of either control siRNA, HO-1 siRNA, NQO1 siRNA, Nrf2 siRNA or HO-1 siRNA plus NQO1 siRNA in combination. A, Protein expression was measured for HO-1 and NQO1 by Western blot analysis. After transfection with siRNA, THP-1 cells were stimulated with LPS for various indicated times and RNA extracts prepared to measure B, HO-1 and NQO1 mRNA expression, C, TNF and IL-1β mRNA expression or D, TNF protein expression. * indicates p<0.05 and ** indicates p<0.01 significance between the different treatment groups.
E, THP-1 cells were transfected with 30 nM of either control siRNA, HO-1 siRNA, NQO1 siRNA or Nrf2 siRNA for 24 h then stimulated with LPS for 30 mins and nuclear extracts prepared. NF-κB DNA binding was assessed using the Panomics p65 NF-κB Transcription Factor ELISA.