Search

Additional file 5: Supplementary Figure 4. Both CDK13-WT and CDK13-Edit are located in the cell nucleus. (A) Schematic representation of the CDK13 domain structure and the CDK13 editing event. Numbers below indicate the amino acid position. Abbreviations: RS: arginine/serine-rich; KD: kinase domain; C: C-terminal extension. (B) Predicted bipartite NLS (Nuclear Localization Sequence) in the WT and c.308 A>G edited form of CDK13. Predictions were performed using NLS mapper ( http://nls-mapper.iab.keio.ac.jp ). (C) Representative western blotting for HA and CDK13 in the indicated Cal62 and TPC1 cells after cytoplasm-nucleus fractionation. SP1 and tubulin were used as loading control for the nucleus and the cytoplasm, respectively.

Additional file 3: Supplementary Figure 2. CDK13 levels in cells stably expressing CDK13-WT, CDK13-Edit or the Empty vector. Cal62 and TPC1 were infected with lentivirus expressing the WT or the edited form of CDK13 (CDK13-WT and CDK13-Edit, respectively) or the corresponding empty vector, and selected using puromycin. (A) CDK13 levels in the indicated cell lines. (B) Representative western blotting for HA antibody in the indicated cell lines. Vinculin was used as loading control (n = 3). Error bars indicate standard deviations. Asterisks denote statistical significance compared with siControl treatment assessed by Student’s t-test (two-tailed). n.s: non significative p > 0.05, ** p < 0.01, *** p < 0.001.

Additional file 1: Table S1. Primers

Additional file 2: Supplementary Figure 1. ADAR1 silencing. Representative western blot of ADAR1 steady-state expression at the indicated time points after ADAR1 silencing in Cal62 and TPC1 cell lines. GAPDH was used as a loading control.

Additional file 4: Supplementary Figure 3. CDK13 overexpression and ADAR1 silencing in Cal62 and TPC1 cell lines. RNA relative levels for CDK13 (A) and ADAR1 (B) in Cal62 and TPC1 cell lines stably expressing CDK13-Edit 72 hours after ADAR1 siRNA (siADAR1 #1 and #2) or Control siRNA (siControl) transfection. (C) Stably-transduced Cal62 and TPC1 cells with CDK13-Edit, CDK13-WT or empty vector were silenced for ADAR1 and were assayed for cell viability by XTT dye reduction (n = 4). Asterisks denote statistical significance assessed by Student’s t-test (two-tailed). n.s: non significative p > 0.05, ** p < 0.01, *** p < 0.001.