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. 2015 May 22;5:9795. doi: 10.1038/srep09795

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Copyright © 2015, Macmillan Publishers Limited

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Functional analyses of survivin-2B-overexpressing RA-FLS.The IVT mRNA (mock and survivin-2B) transfection was used for overexpression experiments for RA-FLS. (a) A Western blot analysis showed that survivin-2B mRNA expressed survivin-2B protein. The cropped blots were used in the figure. (b) The viability of RA-FLS transfected with mRNA (n = 6) at 36hours as determined using the calcein assay. (c,d) Measurement of the Ki-67-positive RA-FLS transfected with mRNA at 48hours (n = 4). The positive rate was calculated as the number of Ki-67-positive cells among the total RA-FLS. (d) FACS plot for mock mRNA (left) and survivin-2B mRNA (right). (e–g) The annexin V and PI analysis for RA-FLS transfected with mRNA at 24hours (n = 4). H₂O₂ stimulation (0.1 mM) was performed for 3hours before the analysis. (e) The percentage of live cells was calculated as the number of annexin V- and PI-double negative cells among the total RA-FLS. (f) The percentage of apoptotic cells was calculated as the number of Annexin V-positive and PI-negative cells among the total RA-FLS. (g) FACS plot for mock mRNA (left) and survivin-2B mRNA (right). The results are the Means ± SEM. t-test ^∗p < 0.05, ^∗∗ p < 0.01.