Characterization of the TCZP-AF4ST stable cell line. A. Stable integrated vectors used in this study. B. Quantitative analysis of relative amounts of AF4 mRNA by qPCR from prepared cDNA with (TCZP+) or without (TCZP-) application of doxycyclin compared to regular 293T cells. Amounts are normalized to Actin [n = 3, +S.E.M.]. C. Top: Representative Western-Blot experiments from TCZP-AF4ST whole cell lysates after 2 passages with or without doxycyclin compared to constitutively expressing constructs pEZP and pEZP-BGH. Faster migrating bands are degradation products of AF4. Bottom: Western-Blot experiments from TCZP-AF4ST whole cell lysates 0-72 h after induction of expression. Lanes 481 and 483 differ in the amount of applied doxycyclin (481 = 1 µg/ml, 483 = 3 µg/ml). D. CCK-8 proliferation assays of induced TCZP-AF4ST cells over a time course of 10 days, compared to non-treated 293T cells [n = 3, ±S.E.M.]. Decline of untransfected cells is due to reaching confluency. E. Representative in-vitro GFP fluorescence microscopy images of TCZP-AF4ST cells during the indicated passages (magnification 100x, 0.5 s. exposure).