Figure 2

Helminth soluble factor–mediated modulation in lipopolysaccharide-induced signaling in microglia. Microglia were pulsed with medium alone, with lipopolysaccharide (LPS) (A) or with helminth soluble factor (HSF) and LPS (B). Equal amounts of lysates from microglia pulsed for the periods of time shown were electrophoretically separated, blotted and probed with antibodies specific for phosphorylated nuclear factor κB (NF-κB) p65 subunit (P-p65), p38, c-Jun N-terminal kinase (P-JNK) and extracellular signal-regulated kinase 1/2 (P-ERK1/2) or acetylated NF-κB p65, whereas antibody against glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. The data shown are representative of three independent experiments. (C) The average pixel intensity of the respective bands at 30 minutes and 60 minutes from three independent experiments was measured. This was done using Quantity One 1-D version 4.6.7 software (Bio-Rad Laboratories). Significant differences were measured by using Student’s t-test, and significant values are denoted by asterisks (*P < 0.05 and ***P < 0.001).