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. 2021 Jun 3;118(23):e2024785118. doi: 10.1073/pnas.2024785118

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Fig. 5. — Many Rhox family members represses LINE1 transposition. (A) Luciferase analysis of LINE1 transposition activity in NIH/3T3 cells cotransfected with the mouse ORFeus LINE1 reporter and vectors expressing the indicated Rhox genes or an empty expression vector as a control (n = 3). *P < 0.05. (B) Luciferase analysis of LINE1 transposition activity in HEK-293T cells cotransfected with the human L1_RP LINE1 reporter and vectors expressing the indicated human RHOX genes or an empty expression vector as a control (n = 4). *P < 0.05. (C) Luciferase analysis of LINE1 transposition activity in TCam-2 cells cotransfected with the human L1_RP LINE1 reporter and vectors expressing the indicated human RHOX genes or an empty expression vector (n = 3). *P < 0.05. (D) Luciferase analysis of LINE1 transposition activity in K562 cells cotransfected with the human L1_RP LINE1 reporter and the RHOXF1 or RHOXF2 shRNA vector or a randomly scrambled shRNA vector (n = 3). *P < 0.05. (E) Luciferase analysis of LINE1 transposition activity in HEK-293T cells cotransfected with the human L1_RP LINE1 reporter and vectors expressing WT or mutated RHOXF2 or an empty expression vector (n = 3). *P < 0.05. (F) Western blot analysis of HEK-293T cells transiently transfected with vectors expressing WT RHOXF2 or the RHOXF2 mutants. β-actin was used for the internal control (n = 2). (G, Left) Luciferase analysis of LINE1 transposition activity in HEK-293T cells cotransfected with the human L1_RP LINE1 reporter and vectors expressing WT or truncated nonfunctional RHOXF2 (c.381dupG). (Right) Luciferase analysis of LINE1 transposition activity in HEK-293T cells cotransfected with the human LINE1 (L1_RP) reporter, a constant amount of WT RHOXF2, and different amount of c.381dupG RHOXF2. Cells cotransfected with the human LINE1 (L1_RP) reporter and an empty expression vector serve as the control (n = 3). *P < 0.05. (H) Western blot analysis of HEK-293T cells transiently transfected with a vector expressing WT RHOXF2 or the RHOXF2 mutants shown. Cells that were transiently transfected with an empty expression vector is the control (n = 2).

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