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. 2020 Jun 29;11(8):2196–2208. doi: 10.1111/1759-7714.13527

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© 2020 The Authors. Thoracic Cancer published by China Lung Oncology Group and John Wiley & Sons Australia, Ltd

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.

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The target relationship between miR‐519d‐3p and HMGB1. (a) The StarBase algorithms showed the complementary binding sequence of miR‐519d‐3p on the wild‐type of HMGB1 3'UTR (HMGB1 3'UTR WT). The mutant type of HMGB1 3'UTR (HMGB1 3'UTR MUT) was constructed. 3'UTR, 3' untranslated region. (b and c) Dual‐luciferase reporter assay evaluated relative luciferase activity of HMGB1 3'UTR WT/MUT in A549 and H1975 cells transfected with miR‐519d‐3p or miR‐NC () miR‐NC, () miR‐519d‐3p; () miR‐NC, ()miR‐519d‐3p. (d and e) Western blotting confirmed the expression of HMGB1 protein in (d) three paired tissues (Normal and Tumor groups), and (e) 16HBE, A549 and H1975 cells. (f) RT‐qPCR detected miR‐519d‐3p expression in A549 and H1975 cells transfected with miR‐519d‐3p or miR‐NC () miR‐NC, () miR‐519d‐3p. (g and h) Western blotting examined HMGB1 protein expression in A549 and H1975 cells transfected with anti‐NC, anti‐miR‐519d‐3p, miR‐NC, or miR‐519d‐3p. (i and j) Western blotting examined HMGB1 protein expression in A549 and H1975 cells transfected with sh‐NC or sh‐circ, and cotransfected with sh‐circ and anti‐NC or anti‐miR‐519d‐3p. **P < 0.01.

Inline graphic — The target relationship between miR‐519d‐3p and HMGB1. (a) The StarBase algorithms showed the complementary binding sequence of miR‐519d‐3p on the wild‐type of HMGB1 3'UTR (HMGB1 3'UTR WT). The mutant type of HMGB1 3'UTR (HMGB1 3'UTR MUT) was constructed. 3'UTR, 3' untranslated region. (b and c) Dual‐luciferase reporter assay evaluated relative luciferase activity of HMGB1 3'UTR WT/MUT in A549 and H1975 cells transfected with miR‐519d‐3p or miR‐NC () miR‐NC, () miR‐519d‐3p; () miR‐NC, ()miR‐519d‐3p. (d and e) Western blotting confirmed the expression of HMGB1 protein in (d) three paired tissues (Normal and Tumor groups), and (e) 16HBE, A549 and H1975 cells. (f) RT‐qPCR detected miR‐519d‐3p expression in A549 and H1975 cells transfected with miR‐519d‐3p or miR‐NC () miR‐NC, () miR‐519d‐3p. (g and h) Western blotting examined HMGB1 protein expression in A549 and H1975 cells transfected with anti‐NC, anti‐miR‐519d‐3p, miR‐NC, or miR‐519d‐3p. (i and j) Western blotting examined HMGB1 protein expression in A549 and H1975 cells transfected with sh‐NC or sh‐circ, and cotransfected with sh‐circ and anti‐NC or anti‐miR‐519d‐3p. **P < 0.01.