COBRA was performed by bisulfite-treating genomic DNA of control and stable Dali knockdown cells, proliferating and differentiated with RA (+RA), PCR-amplifying the CpG-island associated promoters of the indicated Dali-bound and regulated genes, and digesting the PCR products with COBRA-compatible or control enzymes.
For viability, migration, and differentiation analyses, neurospheres were preincubated for 1 week with PBDEs (0.1, 1, or 10 μM) in proliferation medium; afterward, differentiation was initiated and spheres differentiated with the same concentrations of PBDEs in differentiation medium for 48 hr (migration measurements) or 7 days (differentiation analyses).
Finally, we were able to discover meaningful functional networks that were spatially and temporally differentiated with the particular mechanisms varying from trophic associations through interactions with climate and commercial fisheries.
To see whether p53 from stem cells is acetylated, we treated stem cells and differentiated cells with the HDAC inhibitors trichostatin A and nicotinamide prior to cell lysis.
We found that there were no differences in marker expression in spherical and differentiated cells, with the exception of two adhesion molecules, CD44 and CD54.
The medium was changed every other day and cultures were differentiated with the addition of differentiation media when they reached 60 70 % confluency.
Based on the results obtained in earlier studies, malignant and benign spectra were differentiated with the aid of a mathematical algorithm.
Alleles Glu-A3d and Glu-A3g could be differentiated with the aid of the gliadin SDS-PAGE gel; by the presence or absence of the Gli-A1o allele, which we believe is linked to Glu-A3d, but not to Glu-A3g.
