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. Author manuscript; available in PMC: 2015 Jun 26.
Published in final edited form as: J Leukoc Biol. 2006 Mar 16;79(6):1242–1251. doi: 10.1189/jlb.0805456

Fig. 6.

Fig. 6

Effect of SRTK and Syk inhibitors on fibrocyte differentiation. (A) PBMC were incubated for 60 min at 4°C with 10 nM PP2, PP3, or Syk inhibitor. PBMC were then washed twice and cultured in the presence or absence of heat-aggregated, human IgG for 60 min at 4°C. PBMC were then washed twice and cultured at 2.5 × 105 per ml for 5 days in serum-free medium. Results are expressed as the mean ± sem of the number of fibrocytes per 2.5 × 105 cells (n=3 separate donors). Compared with PBMC cultured in SFM, aggregated IgG inhibited fibrocyte differentiation significantly (P<0.01). Compared with PBMC cultured with 10 µg/ml aggregated IgG, PBMC preincubated with PP2 (P<0.05) or the Syk inhibitor (P<0.01) significantly inhibited the ability of IgG to inhibit fibrocyte differentiation as determined by ANOVA. (B) PBMC at 2.5 × 105 per ml were incubated for 60 min at 4°C with 10 nM PP2, PP3, or Syk inhibitor. PBMC were then washed twice and cultured for 5 days in the presence or absence of 0.5 µg/ml SAP. Results are expressed as the mean ± sem of the number of fibrocytes per 2.5 × 105 cells (n=3 separate donors). Compared with PBMC cultured in SFM, SAP inhibited fibrocyte differentiation significantly (P<0.01), as determined by ANOVA. Compared with PBMC cultured in 0.5 µg/ml SAP, preincubation with PP2 significantly inhibited the ability of SAP to prevent fibrocyte differentiation (P<0.01), as determined by ANOVA. (C) PBMC were incubated for 60 min at 4°C with 10 nM PP2, PP3, or Syk inhibitor. PBMC at 2.5 × 105 per ml were then cultured in serum-free medium for 5 days in the presence or absence of 1 µg/ml of the indicated murine F(ab′)2 anti-FcγR antibodies in the presence or absence of 500 ng/ml goat F(ab′)2 anti-mouse IgG. Compared with control cultures, PBMC cultured with anti-FcγRI, anti-FcγRII, or both antibodies inhibited fibrocyte differentiation significantly (P<0.05), as determined by ANOVA. Results are expressed as the mean ± sd of the number of fibrocytes per 2.5 × 105 cells (one of two separate donors). *, P < 0.05; **, P < 0.01.