Interaction between CCN4 and TGF-β3. hBMSCs were transduced with adCCN4 and/or TGF-β3-DDK-over expression vector, and incubated for 2 days, before collection of cell lysates. (A, B) Western blotting was performed using anti-CCN4 antibody or anti-DDK antibody, respectively. (C) Immunoprecipitation assay was performed using an anti-DDK antibody, and CCN4 or TGF-β3/DDK was detected by immunoblotting using an anti-CCN4 or anti-DDK antibodies. (D, E) hBMSCs were transduced with adCCN4 or siCCN4, respectively. After two days, cells were fixed and, the amount of CCN4 bound to cells was measured by its immunoreactivity to anti-CCN4 antibody. (F, G) To analyze the interaction of CCN4 and TGF-β3, fixed cells were incubated with TGF-β3 for 2 h. The amount of TGF-β3 bound to cells was measured by its immunoreactivity using anti-DDK antibody. Bars represent the mean values and standard deviation (+/−SD) (n = 3). **p < 0.01, ***p < 0.001 (Student’s t-tests). Results are representative data of at least three independent experiments.