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. 2020 Sep 17;20(5):250. doi: 10.3892/ol.2020.12113

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Copyright: © Weng et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Figure 1. — Construction of plasmids and evaluation of the knockdown efficiency of TUFM shRNA. (A) pcDNA3.1-EGFP plasmid was used as the vector to construct the pcDNA3.1-TUFM-shRNA plasmid. (B) Following transfection with pcDNA3.1-TUFM-shRNA plasmid, EGFP expression was observed in GIST-T1 (magnification, ×100) and GIST-IR cells (magnification, ×40) by inverted fluorescence microscopy. The results demonstrated that 75.2% of cells expressed the EGFP protein in both cell lines. (C) The results of reverse transcription-quantitative PCR demonstrated that the expression levels of TUFM mRNA in the TUFM shRNA group were lower than those in the control group. (D) Western blot analysis revealed that the protein expression levels of TUFM in GIST-T1 cells transfected with TUFM shRNA plasmid were significantly lower compared with those in the control group. The TUFM level was normalized to the protein level of GAPDH. *P<0.05 and **P<0.01. TUFM, mitochondrial Tu translation elongation factor; shRNA, short hairpin RNA; GIST, gastrointestinal stromal tumor; EGFP, enhanced green fluorescent protein; ctrl, control.