Fig. 3

CXCL12-induced SMAD3 nuclear translocation and recruitment of SMAD3 on the connective tissue growth factor (CTGF) promoter in WI-38 cells. a WI-38 cells were exposed to 10 ng/ml of CXCL12 for 0~ 30 min. In a confocal microscopic image analysis (200×), cells were marked with SMAD3, and nuclei were specified by DAPI. Slides are illustrative of similar results from three separate experiments. b Cells were exposed to 10 ng/ml of CXCL12 for 0~ 30 min. Nuclear protein were prepared and immunodetected with specific antibodies for SMAD3 or lamin A/C. Traces are illustrative of three separate experiments. c Cells were exposed to 10 ng/ml of CXCL12 for 20 min and identified using ChIP assay as defined in “Materials and Methods”. PCR amplification using primers designed against the SMAD3-binding site was performed. Identical volumes of soluble cross-linked chromatins existing in each PCR were approved by the product for input. Negative control: A rabbit polyclonal IgG antibody. Typical traces are illustrative of similar results from three separate experiments