Western blots, showing GFP-aurC and GFP-alone proteins after 24 hours of transient transfection with GFP-alone, GFP-aurC-KD GFP-aurC-CA and GFP-aurC-WT plasmid DNA with mouse Anti-GFP antibody (A) and with rabbit Anti-aurC antibody (B). Western blots showing the level of expression of GFP-aurC protein in three stable clones of GFP-aurC-KD (KD1 to KD3), four stable clones of GFP-aurC-CA (CA1 to CA4), three stable clones each of GFP-aurC-WT (WT1 to WT3) and GFP-alone (GFP1 to GFP3) illustrating the different level of expression of GFP-aurC and GFP proteins by different clones. The antibody used was mouse anti-GFP (C& D) and anti-β tubulin antibody as a loading control (E & F); (G) Kinase assay GFP-aurC-WT, GFP-aurC-CA and GFP-alone clones, using histone-H3 as a substrate. (H 1,2,3,4) The left column shows DAPI stained cells and the right column shows phosphorylated cells with Histone-H3 ser-10. (H-1) GFP-aurC-WT and (H-2) GFP-aurC-CA (H-3) GFP-aurC-KD. (H-4) GFP-alone (I) Histogram shows the percentage of cells with phosphorylation on histone H3 of GFP-aurC-WT, GFP-aurC-CA, GFP-aurC-KD and GFP-alone.
Khan et al. BMC Cell Biology 2012 13:8 doi:10.1186/1471-2121-13-8