Figure 4.

OGT affects MYC-dependent transcription. The expression of MYC was induced in the LNCaP-MYC cell line by addition of doxycycline (Dox). A) Validation of the RNA-seq data using RT-qPCR. Treatment time is 24 hours and OSMI-2 dose 40µM. MMP7, ZNF (=ZNF812P), GINS2, ASF1B, DTL and KLK2 were down-regulated in the combination treatment with OSMI-2 and doxycycline (to induce MYC), while LTF, FBX (=FBXO32) and PRKN were up-regulated. Data shown is an average of three biological replicates with SEM. Student's t-test was used to evaluate statistical significance between Dox and Dox+OSMI-2 condition, *<0.05 and **<0.01. B) Cells were treated with Dox and 40µM OSMI-2 for 24 hours and analyzed by western blotting. C) Pearson correlation of mRNA and protein expression for the proteins that were induced by MYC-overexpression and antagonized by OSMI-2. D) MYC-induced upregulation of PLK1 and Cyclin B1 is antagonized by OSMI-2 (OS2). Cells were treated as indicated and samples were analyzed using western blotting. E) Expression of the LTF gene in prostate cancer patient samples. Data was downloaded from http://www.betastasis.com/prostate_cancer/taylor_et_al_2010. The mean values with SEM are shown for each group. The significance of the data was evaluated using Student's t-test in comparison to normal samples, *<0.05, **<0.01. F) LNCaP-MYC cells were grown in the absence of androgens for 3 days, treated as indicated for an additional 24 hours, protein lysates were collected and western blot was used to detect the indicated proteins.