Figure 4
TSLP increased the expression of CD11b and CD14. THP-1 cells (3 × 105) were treated with IL-32 (0.1 μg/ml), recombinant TSLP (20 ng/ml), or TSLP neutralizing antibodies (1 μg/ml) for 6 days in RPMI1640 containing 10% FBS. (A) Total RNA was assayed by real-time PCR for CD11b and CD14. (B) CD14 was determined by Western blot analysis. THP-1 cells were transfected with TSLP siRNA. Transfected THP-1 cells (3 × 105) were treated with IL-32 (0.1 μg/ml) or recombinant TSLP (20 ng/ml) for 6 days. (C) The total RNA was assayed by RT-PCR analysis for TSLPR and TSLP mRNA. (D) Real-time PCR of macrophage markers, CD11b and CD14 after simulation of THP-1 cells. (E) CD11b (red) and CD14 (green) were examined with a confocal laser scanning microscope. Blank, unstimulated cells; Neu ab, TSLP neutralizing antibodies. *P < 0.05; significantly different from the unstimulated cells' value, **P < 0.05; significantly different from the TSLP-stimulated cells' value. (Original magnification × 600). FBS, fetal bovine serum; IL, interleukin; RT-PCR, reverse transcriptase polymerase chain reaction; TSLP(R), thymic stromal lymphopoietin (receptor).