Regulation of hARD1, hARD2 and NATH during differentiation. NB4 cells were treated with 1 μM all-trans retinoic acid for 96 hours. Untreated wells were cultured in parallel as a negative control. Cells were lysed and analyzed by SDS-PAGE and Western blotting. 10 μg total protein was loaded in each well. Membranes were incubated with the indicated antibodies, anti-hARD1, anti-hARD2, anti-NATH and anti-β-tubulin. The data presented was representative of four independent experiments. Protein levels were quantitated using FUJIFILM IR LAS 1000 and Image Gauge 3.45. Protein levels in control (-) samples were set to 1.0 and protein levels in treated cells (+) were estimated relative to this and normalized to B-tubulin levels. Pictures in the lower panel show representative cells after 96 hours of treatment (+) or control (-). Cells were stained using May-Grünwald-Giemsa.
Arnesen et al. BMC Biochemistry 2006 7:13 doi:10.1186/1471-2091-7-13