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. 2008 Oct 17;179(2):138–150. doi: 10.1164/rccm.200804-535OC

Figure 7.

Figure 7.

Respiratory syncytial virus (RSV)-induced up-regulation and activation of lung Nrf2 in Nrf2+/+ mice. (A) Representative digitized image from semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) demonstrates increased Nrf2 mRNA expression in Nrf2+/+ mice. 18s rRNA bands are shown as matching internal controls. Quantitative RT-PCR assessed the relative ratio of Nrf2 cDNA levels over the vehicle controls after normalization to corresponding 18s rRNA levels. Data are presented as the normalized group mean ± SEM (n = 3/group). *Significantly different from vehicle control mice (P < 0.05). (B) Western blot analysis of lung nuclear proteins from Nrf2+/+ mice determined that RSV exposure increased nuclear translocation of Nrf2 indicated by enhanced intensity of 57-kD bands after RSV. Representative digitized bands from multiple analyses (n = 2/group) are presented. (C) Nuclear protein–antioxidant response element (ARE) binding activity was determined by EMSA using an aliquot of nuclear protein from Nrf2+/+ mice. Increased total ARE-nuclear protein binding activity was determined by enhanced intensity of shifted bands (arrow, left panel). Specific binding activity of small Maf (MafF/G/K), an Nrf2 heterodimer partner for ARE binding, was detected as the appearance of supershifted bands (arrowhead) by adding anti-small Maf antibody in the reaction mixture (+Ab, right panel). Representative images from separate analyses are presented. FP = free probes.