Fig. 5.
A heat-labile protein-like molecule(s) secreted by GR-1 preferentially induces G-CSF production. a Immortalized BMDMs were treated with indicated concentrations of GR-1- or S. aureus-n-butanol-extracted crude LTA. Production of TNF-α (in 4 h) and G-CSF (in 24 h) was measured from spent cell culture media using ELISA. Data shown as mean ± SEM (n ≥ 3; *, p < 0.05 by Student’s t-test). b Immortalized BMDMs were treated with GR-1-cell free spent culture supernatant (SCS; 1/25 dilution) or MRS culture media (1/25 dilution) which had been treated with proteinase K (200 μg/mL), lipase (200 μg/mL), trypsin (200 μg/mL), DNase (200 μg/mL), RNase (200 μg/mL), and heat (95o C for 3 h). Production of TNF-α (in 4 h) and G-CSF (in 24 h) was measured from spent cell culture media using ELISA. Data shown as mean ± SEM (n ≥ 3; *, p < 0.05 by Student’s t-test). c Immortalized BMDMs were treated with MRS culture media (1/25 dilution), GR-1-SCS (1/25 dilution) or GR-1-SCS which had been fractioned using Centricon centrifugal filter devices with a 100 or 30 kDa molecular weight cutoffs. Production of TNF-α (in 4 h) and G-CSF (in 24 h) was measured from spent cell culture media using ELISA. Data shown as mean ± SEM [n ≥ 3]