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. 2017 Jun 27;38(2):829–836. doi: 10.3892/or.2017.5757

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Copyright: © Yang 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 4. — Nicotinic acid (NA) post-transcriptionally downregulates paxillin. U251 cells were treated with the indicated concentration of NA for 4 h. (A-F) DAPI labeling (blue) for nuclei, rhodamine phalloidin labeling for F-actin (red) and immunocytochemistry for p-paxillin (green) were carried out as described in Materials and methods. Regions denoted with rectangles are amplified and shown in insets with red and green channels merged. (G) Western blot analyses for whole-cell lysates were performed with an anti-paxillin antibody. Membranes were stripped and reblotted for p-paxillin and β-actin. (H) Total RNA was extracted, and semi-quantitative RT-PCR was carried out for paxillin mRNA. GAPDH was included as a loading control; -RT, control with no reverse transcriptase (same below). (I) Ectopic expression of 2.5 µg of paxillin in U251 cells, significantly increased paxillin mRNA.