Fig. 6

Anti-fibrotic effects of ENO1 Ab HL217 in TGF-β-stimulated primary human lung fibroblasts. Primary normal human lung fibroblasts (NHLF) (A, C) or diseased human lung fibroblasts from IPF patient (DHLF-IPF) (B, D) were treated with 10 ng/ml TGF-β for 24 h and subjected to the measurement of surface ENO1 expression by using immunofluorescence staining. (A, B) Representative pictures of 4 independent experiments were shown. Cells expressing surface ENO1 were indicated with white arrowhead in the upper panel (ENO1 expression shown in green) and lower panel (merged with nuclear staining DAPI to indicate location of all cells) was kept without arrowhead for clarity. Cells expressing surface ENO1 were manually counted in 5 area of one high power field picture. Four pictures of each group were taken and counted. (C, D) Percentages (%) of surface ENO1 expressing cells was determined over the number of all nucleated cells. NHLF (E, G) and DHLF-IPF (F, H) were treated with 10 ng/ml TGF-β for 4 h and allowed to migrate for another 18 h in the absence or presence of indicated concentrations of HL217, hIgG1, or TXA in the migration assay (E, F) or the CXCL12 chemotaxis assay (G, H). ^#P < 0.05, ^##P < 0.01 vs. untreated group; *P < 0.05, **P < 0.01 vs. TGF-β-treated group (E, F) or TGF-β- and CXCL12-treated group (G, H); ^&P < 0.05 vs. the group with only CXCL12 as chemoattractant. Pooled data of two independent experiments were shown as mean ± SD (one technical replicate and one biological repeat per group for each experiment)