Figure 1From: TIPT2 and geminin interact with basal transcription factors to synergize in transcriptional regulationRNA and protein expression of TIPT2. (A) TIPT2 transcripts in mouse adult organs and embryonic (E14) tissues detected by Northern blotting. Beta actin was used as a loading control. (B) TIPT2 transcript in embryonic tissues prepared at the indicated stages of development detected by RT-PCR. a, m, and p RNAs were prepared from anterior, middle and posterior embryonic regions, respectively. (C) Bacterially made TIPT2 proteins. Lane 1: Marker. Lane 2: Coomassie blue stained recombinant GST-TIPT2. Lane 3: TIPT2, generated by thrombin cleavage, removal of GST by glutathione chromatography, and thrombin removal by benzamidine chromatography. Lane 4: Material equivalent to the protein in lane 3 was cut from a gel and electrophoresed again. (D) TIPT2 proteins detected by purified anti-TIPT antibodies in HeLa or U2OS whole cell extracts (lane 2 and 3). Detection of TIPT2 on the Western blot was competed by the inclusion of GST-TIPT2 in the antibody binding solution in a 10:1 ratio (lane 4). Markers were run on lanes 1 and 5. (E) TIPT2 proteins in organ extracts from P2 mice, adult mouse testis and a stable U2OS cell line expressing FHM-TIPT2. Asterisks indicate additional proteins recognized by anti-TIPT2 antibodies. (F) Cytoplasmic, nucleoplasmic and nucleolar localization of TIPT2 in human U2OS tissue culture cells. Nucleoli were detected with anti-TBPL1 antibodies. DNA was stained with DAPI. Bar, 20 μm. Further immunohistochemistry is presented in Additional file 1.Back to article page