Additional file 9: Figure S8. of Knockdown of NAT12/NAA30 reduces tumorigenic features of glioblastoma-initiating cells

Expression of the selected genes and proteins in the NAT12/NAA30 knockdown cultures. A, Western blot showing that the protein levels of active β-Catenin (ABC), phospho-p70 S6 kinase (Thr 389), phospho-STAT3 (Ser727) and GLI1 were slightly affected or not altered in the knockdown cultures. The signals obtained with Anti-SHH and Anti-phospho-p53 (Ser15) were below the detection level of western blot (not shown). For a complete list of ABs see Supplementary File 4. B, Western blots showing the expression of neural stem cell markers SOX2 and NESTIN, proteins representing hypoxia response (KCNMA1), cell cycle (CCND1) and ACTB (ACTIN). C, Western blots showing the expression of p-AKT (Ser473), total AKT, p-AKT (Ser308), p-p4E-BP1 and total p4E-BP1. D, Expression of selected stem cell genes SOX2, NANOG, POU5F1/OCT4, MYC and CCND1 on microarrays (n=4 for each cell culture). None of the selected genes were differentially regulated in the NAT12/NAA30 knockdown cultures. The bottom and top of each box indicate the 25th and 75th percentile (the lower and upper quartiles, respectively), and the band near the middle of the box represents the 50th percentile (the median). Interestingly, the knockdown strength was in correlation with the effect on downstream genes. In the KD2 culture that exhibited stronger knockdown effect at the protein level and better reduction in cell viability and number of spheres we could also detect decreased levels of GLI1, nestin and phospho proteins such as phospho-AKT (Ser308) and p-4E-BP1(Thr37/46) (Figure S8 A-C)