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. 2014 Mar 20;289(19):13531–13542. doi: 10.1074/jbc.M113.525287

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© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — Expression of PTX3 mRNA and protein by AMEC and AMSC. Total RNAs and proteins were extracted from HSF, AMEC, and AMSC cultured in DMEM/F12 with 0.5% FBS for 48 h before being treated with 20 ng/ml of TNF or IL-1β for 4 h (real-time PCR) or 24 h (Western blot analysis). A, the expression of PTX3 mRNA was analyzed by real-time PCR with GAPDH as the endogenous control. *, p < 0.01, statistically significant groups in comparison with the control. B, the expression of PTX3 protein in medium (top panel) and cell lysates (center panel) was compared by Western blot with anti-PTX3 antibody and normalized by the β-actin control. M, protein ladder markers. C, PTX3 siRNA transfection was performed in AMEC and AMSC with or without 20 ng/ml of TNF after being cultured in DMEM/F12 with 0.5% FBS for 48 h. The down-regulation of PTX3 in both medium (top panel) and cell lysates (center panel) by PTX3 siRNA was verified by Western blot analysis. scRNA, scrambled RNA; M, protein ladder markers.