Effects of RPR crude extract on the expression levels of cytokines in BCG-stimulated PBMac. PBMac (5 × 105) were pretreated with 0.1% DMSO or 100 μg/mL RPR extract overnight and then stimulated with BCG (MOI = 1). (A) Total mRNA was extracted after 3 hours for cDNA synthesis. The levels of mRNA were determined by q-PCR. The mRNA level of the DMSO + BCG sample was set as 100%, and the rest were compared to the DMSO + BCG sample to obtain a percentage value. (B) Culture supernatants were collected after 24 hours. The levels of IL-6, IL-8, IL-10 and TNF-α in the supernatants were measured by ELISA. The individual cytokine level in the supernatant of the DMSO + BCG sample was set as 100%, and the rest were compared to it to obtain a percentage value. Results are shown as mean ± SD from independent experiments on PBMac obtained from three (A) or six (B) different healthy donors. *P < 0.05, **P < 0.001 compared to the DMSO + BCG sample (one-way ANOVA, Tukey's test).