Figure 5
Inhibitory effect of CILP1 on TGFβ1 signalling. (A) Inhibitory effect of recombinant CILP1 on TGFβ1-induced ACTA2 overexpression in adult rat CFB (treatment 24 hrs, data from 6 separate cell isolations). (B) Inhibitory effect of recombinant CILP1 on TGFβ1-induced CTGF overexpression in adult rat CFB (treatment 24 hrs, data from 6 separate cell isolations). Data from A en B were normalized to the housekeeping gene Cyclophilin-A using the comparative threshold cycle (Ct) method by calculating 2ΔCt and multiplied by 1000 (formula 1000 * 2ΔCt), to enhance readability. (C) Relative αSMA protein level as determined by ELISA in adult rat CFB treated for 48 hrs with TGFβ1, CILP1 or the combination (data are shown relative to control from 6 separate cell isolations). (D) typical examples of αSMA staining in adult rat CFB incubated for 48 hrs with TGFβ1, CILP1 or the combination. (E) Effect of CILP1 on TGFβ-signalling using a TGFβ promoter/reporter assay. HEK293 cells were cotransfected with CILP1 and the TGFβ promoter/reporter construct, containing SMAD-binding elements (CAGA-Luc) and incubated with TGFβ1 for 24 h before analysis of SMAD-binding activity (data are shown relative to control from 5 separate experiments). All data are expressed as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001.
