Loss of full-length TUT4 and TUT7 in the TUT4/7 double mutants has a negative impact on cancer cell properties. Molecular masses of TUT4, TUT7, and the loading control α-tubulin (TBA4A) are 185, 171, and 50 kDa, respectively. (A,B) Western blot images of TUT4 (A) and TUT7 (B) in independent TUT4/7 double-mutant (cKO) clones (TUT4/7 cKO #1 and TUT4/7 cKO #2) of DU145 (n = 3). (C,D) Western blot images of TUT4 (C) and TUT7 (D) in independent TUT4/7 double-mutant (cKO) clones (TUT4/7 cKO #1 and TUT4/7 cKO #2) of IGROV1 (n = 3). (E,F) Quantitative cell proliferation assay at 2-h intervals using the Incucyte systems. Solid line indicates curve fitted to the logistic equation with r as the growth rate and p as the P-value of r. WT is the control for DU145 (E) and IGROV1 (F). TUT4/7 cKO #1 and TUT4/7 cKO #2 are the TUT4/7 double mutants. The y-axis represents relative percent confluency while the x-axis denotes time in hours (n = 2). (G) Quantitative wound healing assays with measurements at 2-h intervals. The y-axis denotes the wound width in µm and the x-axis represents time in hours. Cell line = IGROV1. WT is the control IGROV1 cell line. TUT4/7 cKO #1 and TUT4/7 cKO #2 are the TUT4/7 double mutants (n = 2).
