Figure 5.

EZH2 and G9a inhibition and knockdown restore CXCL10 expression in F-IPF. F-IPF cells were (A and B) incubated with DZNep (10 nM) or (C and D) transfected with empty vector, pMSCV-mJMJD3, or pMSCV-JMJD3 or (E and F) transfected with control siRNA, EZH2 siRNA, or G9a siRNA in culture medium for 48 hours and in serum-free medium for 24 hours before being treated with IL-1β (1 ng/ml) for a further (A, C, and E) 4 hours or (B, D, and F) 24 hours. Total RNA was isolated, and mRNA levels of CXCL10 and the internal control β2M were determined by real-time RT-PCR. (A, C, and E) Results are calculated as the ratio of CXCL10 mRNA and β2M mRNA. (B, D, and F) Medium was collected, and CXCL10 concentration was analyzed by ELISA. Data are expressed as mean ± SEM of six separate experiments performed in duplicate. *P < 0.05, **P < 0.01 compared with corresponding untreated cells.