Fig. 3

DHT and androstenedione, similarly to DHEA, affect RACK1 expression. a, b Increased RACK1 expression following treatment with DHT and androstenedione. THP-1 cells were treated with increasing concentrations of DHT (1–10 nM), androstenedione (10 nM) or DMSO as vehicle control for 16 h for mRNA expression (a) or 24 h for protein expression (b). RACK1 mRNA expression was assessed by Real Time-PCR. Results are expressed as mean ± SD of three independent experiments, with *p < 0.05 and **p < 0.01 vs Control cells. RACK1 immunoreactivity was assessed by Western blot analysis of cell lysates (b). A representative Western blot of RACK1 immunoreactivity in cell lysate is shown. c, d Effect of DHT and androstenedione on LPS-induced cytokine production. THP–1 cells were treated with increasing concentrations of DHT (1–10 nM), androstenedione (10 nM) or DMSO as vehicle control. After 24 h, LPS (10 ng/ml) was added for 3 h to assess TNF-α release (c) or 24 h to assess IL–8 release (d). Results are expressed as mean ± SD, n = 3 replicates. Data is representative of three independent experiments. Statistical analysis was performed with Tukey’s multiple comparison test with §p < 0.05 and §§p < 0.01 vs LPS treated cells