Fig. 4

Expression of RanBP17 during the cell cycle. A FACS sorting yields 4 cell populations, representing the G1/G0, S and G2/M phases of the cell cycle as well as a population labeled „All“, which contains all phases for comparison. Western blot analysis points to increased expression levels of RanBP17 (antibody orb226830) during the S-phase of the cell cycle, which is particularly prominent in UT-SCC-26A. Cell cycle associated proteins (PCNA, cyclin-B1, CDK1) show an expected increase of their expression during S- and into G2/M-phase, thereby helping to validate the accuracy of cell sorting to the respective phase of the cell cycle. B Total cellular protein was significantly elevated in S phase cells compared with cells in the G1/G0 phase of the cell cycle, which is an expected finding for cells in division. A similar observation was made for total RNA levels. C No significant differences in RanBP17 RNA levels could be noted between the different cell cycle phases for UM-SCC-3 (n=4) and UT-SCC-26A (n=3) cells. Statistical tests: B and C: One way ANOVA with Dunn´s correction for multiple comparisons. *p<0.05. (A: Protein bands shown in the upper two panels (RanBP17-orb226830 and GAPDH) and the lower six panels (RanBP17-GTX70420, PCNA, Cyclin-B1, GAPDH, CDK1, Beta-Tubulin) are derived from lysates of two independent experiments). TP53 mutation in UM-SCC-3: p.R248Q (nuclear p53); TP53 mutation in UT-SCC-26A: p.Y236* (cytoplasmic p53)