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. 2009 Apr 7;30(7):1082–1088. doi: 10.1093/carcin/bgp078

Fig. 3.

Fig. 3.

ANXA1 regulates IL-6 expression in prostatic epithelial cells. (A, C and D) Secreted IL-6 levels were measured from culture supernatants of RWPE-1/NC and RWPE-1/rA1 cells by enzyme-linked immunosorbent assay and data represent the mean ± SD. (graphs). (A) IL-6 secretion was measured from unstimulated RWPE-1/NC and rA1 cells (−) or cells stimulated with 25 ng/ml EGF or 10 ng/ml TNF-α. Unstimulated IL-6 secretion from RWPE-1/rA1 was significantly different from unstimulated RWPE-1/NC by a Student's t-test, **P = 0.0016. (B) Reduction of ANXA1 protein expression in DU145-expressing ANXA1 shRNA and controls was measured by western blotting. (C) IL-6 secretion from DU145 variants was determined by enzyme-linked immunosorbent assay (top) and IL-6 mRNA was estimated by semiquantitative reverse transcription (RT)–PCR (bottom). Double asterisks indicate significant difference from control DU145 cells as determined by Tukey–Kramer multiple comparison test (P < 0.001). (D) Restoration of ANXA1 protein expression suppresses IL-6 secretion. DU145-rA1.7 cells were transiently transfected with yellow fluorescent protein (YFP) or shRNA-insensitive ANXA1-YFP (srA1) expression plasmids and secreted IL-6 levels were measured after 24 h in culture.