Figure 2.
Effect of p402, p40, interleukin-23 (IL-23) and IL-12p70 on the expression of IL-7 in mouse primary macrophages, RAW cells, human and mouse primary astrocytes. Cells were stimulated with different concentrations (5 and 10 ng/ml) of p402, p40, IL-12p70 and IL-23 separately under the serum-free condition. After 4 hr of stimulation, the mRNA expression of IL-7 was monitored by semi-quantitative RT-PCR (a and e) and real-time PCR (b and f). Results are means ± SD of three different experiments.*P < 0·001 versus control IL-7 mRNA. NS, non-significant. After 18 hr of stimulation by 10 ng/ml p402, IL-12p70 and IL-23, the expression of IL-7 protein was monitored by immunofluorescence in peritoneal macrophages (c), RAW cells (d), mouse astrocytes (g) and human astrocytes (h). DAPI was used to visualize the nucleus. Results represent three independent experiments. Mouse primary microglia and astrocytes (5 × 105) were stimulated with IL-12p70, p402 and IL-23 under serum-free conditions. After 12 hr, supernatants were collected. Concentration of IL-7 was measured in culture supernatants by a high-sensitivity ELISA (i). Two separate wells from each treatment group have been processed and shown. Results represent three independent experiments. *P < 0·001 versus control; **P < 0·001 versus control.