Figure 1. Activation of MAPK signalling pathways by CTGF in HMC.

(A) Cells were grown on coverslips and serum-starved for 48 h prior to incubation in the absence or presence of 20 ng/ml r-CTGF for 5 or 30 min. Cells were fixed, permeabilized, probed with anti-phospho-ERK1/2, JNK and p38 MAPK primary antibodies, and then with fluorescein-conjugated secondary antibody, as described in the Materials and methods section. Results are representative of three separate experiments. (B) Serum-starved HMCs were incubated in the presence of r-CTGF for the periods of time indicated, after which the cells were lysed. Equal amounts of lysate protein were subjected to SDS/PAGE and analysed by Western blotting using phospho-specific antibodies against the p38 MAPK and the phosphokinases upstream of it, MKK3/6, as well as antibody against the total p38 MAPK. Results are representative of three separate experiments