Figure 2.

Sustained Inflammation and Activation of Pro-oncogenic Signaling Pathways in LFA- and LNT-Induced Lesions at 6 Months Post-injection

(A) Cell types in LFA- and LNT-induced inflammatory lesions at 6 months post-injection were quantified by immunostaining with a panel of cell markers (pan-cytokeratin, mesothelial cells; CD68, macrophages; CD45, leukocytes; CD3, T cells; Ly6g, granulocytes).

(B) Specific areas of tissue were isolated from fresh frozen diaphragms (n = 4) by laser microdissection in order to examine gene expression levels in different cell types (muscle, mesothelium, and lesion). For validation of the cell-type selection, the expression level of the cell markers mesothelin and CD68 in the different areas sampled was measured by qPCR. The expression level of the genes encoding STAT3, IL-6, and PI3K was examined by qPCR in muscle, mesothelium, and lesion (where present) microdissected from mice exposed to VC, LFA, or LNT at 12 weeks post-injection. ^∗p < 0.05, ^∗∗p < 0.01.

(C) Activation of signaling pathways in the chest wall tissue of mice 6 months post-injection was analyzed by immunostaining. Positive staining for signaling proteins was observed in both mesothelial and non-mesothelial cells (black arrows). Scale bar, 20 μm.

See also Figure S4.