Figure 6.
OBF1 is required for the proliferation of GC-derived lymphoma cell lines. (A) Left, immunoblot showing OBF1 knockdown efficiency using shRNA specific for OBF1 (OBF1-sh1) or control (SHC002); histone 3 (H3) is shown as loading control. Right, assessment of proliferation of Raji cell line following shRNA-mediated depletion of OBF1. Cells were seeded at day 1 and counted every second day until day 7 (n = 3). (B) Left, immunoblot of OCT1 demonstrating knockdown efficiency using shRNA specific for OCT1; actin is shown as loading control. Right, assessment of proliferation of Raji cell line following shRNA-mediated depletion of OCT1 (n = 3). (C) Left, immunoblot of OCT2 demonstrating knockdown efficiency using shRNA specific for OCT2; actin is shown as loading control. Right, assessment of proliferation of Raji cell line following shRNA-mediated depletion of OCT2 (n = 3). (D) Left, immunoblot demonstrating knockdown efficiency using shRNA specific for OCT1 and ectopic expression of OBF1 tagged with AviTag and FLAG; actin is shown as loading control. Right, assessment of proliferation of Raji cell line ectopically expressing AviTag-FLAG tagged OBF1 following shRNA-mediated depletion of OCT1 (n = 3). (E) Left, immunoblot demonstrating knockdown efficiency using shRNA specific for OCT2 and ectopic expression of OBF1 tagged with AviTag and FLAG; actin is shown as loading control. Right, assessment of proliferation of Raji cell line ectopically expressing AviTag-FLAG tagged OBF1 following shRNA-mediated depletion of OCT1 (n = 3). (F) Left, immunoblot showing the downregulation of BCL6 upon shRNA-mediated knockdown of OBF1; MCM7 is shown as loading control. Right, qPCR measurements of BCL6 expression upon OBF1 knockdown (same data as shown in supplemental Figure 5E). (G) Left, confirmation of downregulation of OBF1 by shRNA specific for OBF1 by qPCR (n = 4); right, confirmation of downregulation of IRF4 by shRNA specific for IRF4 by qPCR (n = 4). (H) Assessment of Raji cells proliferation following shRNA-mediated depletion of OBF1 and depletion of both OBF1 and IRF4 (n = 3). (I) Assessment of proliferation of Raji cell line following ectopic expression of IRF4. Cells were seeded at day 1 and counted every second day until day 7 (n = 3). Mean ± standard deviation; *P < .05; **P < .01; ***P < .001; ****P < .0001 (2-tailed Student t test).

OBF1 is required for the proliferation of GC-derived lymphoma cell lines. (A) Left, immunoblot showing OBF1 knockdown efficiency using shRNA specific for OBF1 (OBF1-sh1) or control (SHC002); histone 3 (H3) is shown as loading control. Right, assessment of proliferation of Raji cell line following shRNA-mediated depletion of OBF1. Cells were seeded at day 1 and counted every second day until day 7 (n = 3). (B) Left, immunoblot of OCT1 demonstrating knockdown efficiency using shRNA specific for OCT1; actin is shown as loading control. Right, assessment of proliferation of Raji cell line following shRNA-mediated depletion of OCT1 (n = 3). (C) Left, immunoblot of OCT2 demonstrating knockdown efficiency using shRNA specific for OCT2; actin is shown as loading control. Right, assessment of proliferation of Raji cell line following shRNA-mediated depletion of OCT2 (n = 3). (D) Left, immunoblot demonstrating knockdown efficiency using shRNA specific for OCT1 and ectopic expression of OBF1 tagged with AviTag and FLAG; actin is shown as loading control. Right, assessment of proliferation of Raji cell line ectopically expressing AviTag-FLAG tagged OBF1 following shRNA-mediated depletion of OCT1 (n = 3). (E) Left, immunoblot demonstrating knockdown efficiency using shRNA specific for OCT2 and ectopic expression of OBF1 tagged with AviTag and FLAG; actin is shown as loading control. Right, assessment of proliferation of Raji cell line ectopically expressing AviTag-FLAG tagged OBF1 following shRNA-mediated depletion of OCT1 (n = 3). (F) Left, immunoblot showing the downregulation of BCL6 upon shRNA-mediated knockdown of OBF1; MCM7 is shown as loading control. Right, qPCR measurements of BCL6 expression upon OBF1 knockdown (same data as shown in supplemental Figure 5E). (G) Left, confirmation of downregulation of OBF1 by shRNA specific for OBF1 by qPCR (n = 4); right, confirmation of downregulation of IRF4 by shRNA specific for IRF4 by qPCR (n = 4). (H) Assessment of Raji cells proliferation following shRNA-mediated depletion of OBF1 and depletion of both OBF1 and IRF4 (n = 3). (I) Assessment of proliferation of Raji cell line following ectopic expression of IRF4. Cells were seeded at day 1 and counted every second day until day 7 (n = 3). Mean ± standard deviation; *P < .05; **P < .01; ***P < .001; ****P < .0001 (2-tailed Student t test).

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