Western blot analysis of extracts from various cell lines using PHC1 (1F3F3) Mouse mAb (upper) and GAPDH (D16H11) XP ® Rabbit mAb #5174 (lower). As expected, PHC1 protein is not expressed in DU 145 or NIH/3T3 cells.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 10 6 NCCIT cells and either 10 µl of PHC1 (1F3F3) Mouse 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 SimpleChIP ® Human HoxA1 Intron 1 Primers #7707, SimpleChIP ® Human HoxA2 Promoter Primers #5517, and SimpleChIP ® Human α Satellite Repeat Primers #4486. 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.