Figure 3

OSM and other gp130 cytokine stimulation and HASMC responses. (A-C) HASMC cultures were prepared and stimulated as in Figure 1 with the indicated cytokines and concentrations of 10 ng/ml for each. Data shown are from one of two individual cell lines that showed identical trends. 24-hour culture supernatants were assessed by ELISA for levels of MCP-1/CCL-2 (A), IL-6 (B) and eotaxin-1 (C). ***p < 0.001 indicates statistical significance comparing indicated cytokine combinations to either cytokine alone using One-Way ANOVA with Bonferroni’s post-test. (D) HASMC cultures were stimulated with OSM, LIF, IL-11, IL-31 or IL-6 (10 ng/ml) for 20 minutes and cell lysates were prepared for immunoblots as described in methods. The lysates were probed for p-Y-STAT1, STAT1, p-Y-STAT3, STAT3, p-Y- STAT5, STAT5, p-Y-STAT6, p-Ser-Akt, p-T/Y-p38, p-T/Y-JNK and Actin as indicated. (E) Quantitative analysis of band intensity using densitometry (ImageJ) corrected to Actin or total protein for each probe and expressed as fold change relative to control (unstimulated).