Western blot analysis of extracts from MEF wt and U-2 OS cells, untreated (-) or treated with MG-132 #2194 (10 μM, 10 hr; +), using NRF2 (D1Z9C) XP ® Rabbit mAb.
Immunoprecipitation of NRF2 from MEF wt cell extracts treated with MG-132 #2194 (10 μM, 10 hr) using Rabbit (DA1E) mAb IgG XP ® Isotype Control #3900 (lane 2) or NRF2 (D1Z9C) XP ® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using NRF2 (D1Z9C) XP ® Rabbit mAb (lane 3).
Flow cytometric analysis of MEF wt cells, untreated (blue) and treated with MG-132 #2194 (green), using NRF2 (D1Z9C) XP ® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab') 2 Fragment (Alexa Fluor ® 647 Conjugate) #4414 was used as a secondary antibody.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 MEF NRF2 wild-type (left) and NRF2 knock-out (right) cells, both treated with DEM (50 μM, 3 hr), and 5 µl of NRF2 (D1Z9C) XP ® Rabbit mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP ® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using mouse MafG intron 1 primers, SimpleChIP ® Mouse NQO1 Promoter Primers #12635, and SimpleChIP ® Mouse RPL30 Intron 2 Primers #7015. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.