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. 2023 Jul 9;25(11):2015–2027. doi: 10.1093/neuonc/noad111

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Figure 5. — E2F transcription factor 1 (E2F1) directly activates the transcription of nonstructural maintenance of chromatin condensin I complex subunit G (NCAPG). (A, B) The protein and mRNA expression of NCAPG and E2F1 were detected. (C) The correlation of NCAPG and E2F1 expression was analyzed. (D) The Dual-Luciferase reporting gene assay was performed to investigate the effect of E2F1 overexpression or E2F1 knockdown on NCAPG promoter activity respectively. (E) Predicted sequences of E2F1 motif in NCAPG promoter by JASPAR. (F) The ChIP-sequencing data of E2F1 were downloaded from the Cistrome DB database and visualized by IGV software. (G) ChIP analysis of the region of E2F1 enrichment at NCAPG promoter. (H) Luciferase reporter constructs containing the truncated wild NCAPG promoter or E2F1 motif mutant promoter were con-transfected with a shE2F1 or with an empty vector into 293FT cells, and luciferase activity was evaluated. (I) The Dual-Luciferase reporter gene assay demonstrates the promoter activity of NCAPG in the 293FT cells transfected with His-E2F1 and/or sh polymerase 1 (PARP1). (J) ChIP analysis of E2F1 enrichment at NCAPG promoter in glioblastoma-3 cells expression Scramble or shPARP1. (K, L) The mRNA level of NCAPG in PARP-silenced cells were determined by qPCR assay. All data are shown as the means ± SD; *P < .05, **P < .01, ***P < .001.