Figure EV3. In vitro and in vivo characterization of the early passage cell lines.

• A
  Kaplan–Meier curve of C57BL/6 mice transplanted with the CX-5461‐resistant CX #39 early passage Eμ‐Myc lymphoma cells treated with CX‐5461 (35 mg/kg every 3 days; n = 5–6). Survival curves of drug‐naïve cells were actual data reproduced from Devlin et al, 2016.
• B
  Kaplan–Meier curve of C57BL/6 mice transplanted with CX‐5461 + EV‐resistant CMB #9 early passage cells treated with CX‐5461 (35 mg/kg every 3 days) and everolimus (5 mg/kg daily; n = 5–6). Survival curves of drug‐naïve cells were actual data reproduced from Devlin et al, 2016.
• C
  Western blot analysis of markers for the on‐target effect of 20 nM everolimus treatment for 3 h, specifically phosphorylation of RPS6 at Serine 240/244 residues and total RPS6 (n = 3). Actin was used as a loading control.
• D–J
  Synthesis rates of 47S pre‐rRNAs in (D) drug‐naïve (CTRL), (E) everolimus‐resistant (EV), (F) CX-5461‐resistant (CX), and (G) CX‐5461 + EV‐resistant (CMB) cell lines in response to 3‐h 50 nM CX‐5461 treatment were determined by ³²P‐orthophosphate “pulse” labeling. Synthesis rate of 47S pre‐rRNAs in (H) CX and (I) CMB cell lines in response to 10‐h 50 nM CX‐5461 was determined by ³²P‐orthophosphate “pulse” labeling and (J) its quantitation (n = 3). (D–I) 28S rRNA abundance was used as a loading control. EtBr: ethidium bromide. Data were analyzed by Student's t‐test. Graphs represent mean ± SEM of n = 3. *P ≤ 0.05. ***P ≤ 0.001.
• K
  Western blot analysis for p53 abundance in response to 50 nM CX‐5461 treatment for 3 h. Actin was used as a loading control (n = 2–3).

Source data are available online for this figure.