Fig. 2. In vitro and in vivo effects of enforced KDM5B expression.
a MTT assays with Cpd1 started at day −7, and then MAPKi was added for 72 h. Readout after 72 h of MAPKi (5 µM PLX4720 + 0.5 µM GDC-0973) and in total 10 days of Cpd1. Cell numbers after 10 days of DMSO or Cpd1 without MAPKi (left panel) and after 72 h of MAPKi compared to “no MAPKi control” (right). Mean ± SD (n = 3), two-sided t-test (****p ≤ 0.0001). b Clonogenic growth after KDM5B induction (9, 16, and 20 days). Mean ± SD, two-sided t-test. Shown is one out of six clones. c Soft agar colony formation after 52 days of KDM5B induction. Shown is one representative experiment (n = 3). Mean ± SD; two-sided t-test. d Growth of xenografted WM3734Tet3G-KDM5B cells. Doxycycline treatment started day 34. Mean ± SEM with n = 6 mice in the control and n = 10 mice in the treatment group, linear mixed-effect spline model. e Clonogenic growth assay of WM983B and WM1366 cells continuously treated over 7 and 9 days with 10 µM of Cpd1 or Neg4 (n = 3). f Clonogenic growth assay of 451Lu and resistant 451Lu BR cells treated over 9, 16, and 20 days with 10 µM of Cpd1 vs. DMSO or Neg4 controls (n = 2). g Two-dimensional colony formation after continuous treatment with 10 µM Cpd1 for 19 days. Mean, n = 3. h Soft agar colony formation under constant Cpd1 treatment at the indicated doses over 30 days (one representative experiment out of n = 2) and i compared to short-term treatment for 72 h before seeding without treatment continuation (mean, n = 2). j Collagen invasion of WM3734 spheroids under 10 µM of Neg4 or Cpd1 at day 10. Shown is one representative experiment (n = 3). Mean ± SD; two-sided t-test. k WM3734 melanoma spheroids at day 10 after collagen embedding. l, m Growth of xenografted WM3734 cells (left, n = 9 mice per group) and syngeneic CM cells (right, n = 3 per control and n = 5 mice per treatment group). Cpd1 treatment was started when tumors were palpable. Mean ± SEM, linear mixed-effect spline model. Source data are provided as a Source Data file.