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. 2012 Nov 8;8(11):e1003048. doi: 10.1371/journal.pgen.1003048

Figure 4. Oct1 RNAi diminishes the side population in A549 cells.

Figure 4

A. SP assay of luciferase-positive A549 cells carrying an inducible scrambled or Oct1-specific shRNA [21]. Cells were treated with doxycycline for 4 days. Side population assays were conducted as described in the methods section. B. Western blot showing Oct1 protein levels in the two cell lines from (A) with and without 4-day culture in doxycycline. ß-actin is shown as a loading control. C. Quantification of the changed in SP percentage from three independent trials. Error bars denote standard deviations. D. Alignment of Abcg2 first intron regions in human and mouse. The perfect octamer sequence in both species is highlighted. Human Abcg2 has two annotated transcription start sites, so the element is located at both +14 kb and −12.4 kb relative to the transcription start sites. E. Quantification of ChIP enrichment using A549 cells and specific antibodies directed against Oct1, Mta2 and Jmjd1a. The PCR primer pair spanned the human Oct1 binding site in Abcg2. ChIP enrichment was quantified relative to isotype control anti-C/EBPß antibodies and relative to a control region as described in the methods section. Values are the average of four independent experiments. Error bars represent standard deviations.