JCS03310 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Suppression of CD44-HA interaction by a neutralizing anti-CD44 antibody. (A) HEK293 cells were transfected with a control (mock) or a CD44 plasmid. Cell surface levels of CD44 were analyzed by flow cytometry. (B) The mock- and CD44-transfected cells were incubated with a control Ig or a neutralizing anti-CD44 Ab. Cells were then stained with FITC-conjugated HA and subjected to flow cytometry. The black lines show cells stained with FITC secondary Ig conjugate.

• Supplemental Figure 2 -

  Fig. S2. Expression of TLR4 on BMMs. BMMs were incubated with TLR4 Ab followed by FITC-conjugated secondary Ab incubation. Cells were analyzed by a flow cytometer.

• Supplemental Figure 3 -

  Fig. S3. Differential effects of HA from effects of LPS. BMMs were primed with M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 3 days. The cells were then treated with LPS (0.001-1 ng/ml) or HA (1-5 μg/ml) in the presence of M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 16 hours. Cells were stained for TRAP and multinuclear TRAP⁺ cells were counted. *P<0.05, **P<0.01 as compared with vehicle-treated control.

• Supplemental Figure 4 -

  Fig. S4. No effects of HA on the surface level of M-CSF receptor. BMMs were cultured with M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 2 days in the absence or presence of HA (2 μg/ml). Cell surface expression of c-Fms, the M-CSF receptor, was analyzed by flow cytometry. Blue line, −HA; red line, +HA; black area, secondary Ab control.