Effects of RPR-EA-S1 on signaling kinases and NF-κB expression in BCG-stimulated PBMac. PBMac (2 × 106) were pretreated with 0.01% DMSO or 20 μg/mL of RPR-EA-S1 overnight and then stimulated with BCG (MOI = 1) for 30 minutes. Cytoplasmic proteins and nuclear proteins were extracted separately. (A) and (C): The levels of phospho-MAPK, phospho-GSK and phospho-Akt, as well as NF-κB1 p50 and IκBα were analyzed with Western blot. The figures showed one set of representative results from independent experiments on PBMac obtained from three different healthy donors. (B) and (D): Quantification of the Western blot was measured by laser densitometry. Intensities of phospho-MAPK, phospho-GSK, phospho-Akt, NF-κB1 p50 and IκBα were normalized to the corresponding MAPK, GSK, Akt, Lamin B and Actin, respectively. Results are shown as mean ± SD from independent experiments on PBMac obtained from three different healthy donors. *P < 0.05 compared to the DMSO + BCG sample (one-way ANOVA, Tukey's test).