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. 2020 Sep 8;10:1730. doi: 10.3389/fonc.2020.01730

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Copyright © 2020 Li, Ge, Chen, Sun, Jiang, Xie, Lu, Zhang, Han, Chen and Zhu.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Expression analysis for genes that as the G1-S phase and DNA replication regulators after RPL21 knockdown. (A,B) The validation of transcriptome analysis by quantitative real-time PCR (qPCR). Human pancreatic cancer PANC-1 cells were transfected with siL21-Mix (40 nM, 72 h), the up-regulated genes (AHR, THBS1, DDIT3, and MKNK2) in transcriptome sequencing were confirmed by qPCR. The down-regulated genes (E2F1, PCNA, CCND1, CCNE1 MCM2, MCM4, MCM5, MCM7, and KIAA0101) in transcriptome sequencing were confirmed by qPCR. (C) The PANC-1 and BxPC-3 cells were transfected with Mock-siRNA, siL21-1 and siL21-2 (40 nM, 72 h). The equal amounts (15 μg) of each protein sample were analyzed by western blot with antibodies of E2F1, CCND1, CCNE1, MCM2-7 and GAPDH. The GAPDH served as an internal control. The control, negative control (NC), siL21-1 and siL21-2 represented the untransfected, Mock-siRNA transfected, siL21-1 and siL21-2 transfected, respectively. Three independent experiments were of similar results.