Expression of extracellular matrix proteins and migration-related genes in HaCaT cells treated with DK223. (A) mRNA expression was examined in HaCaT cells treated with 2 μM DK223 for 24 h using RT-PCR with the specific primers indicated; (B) Cells were treated with 2 μM DK223 in serum-free media. Conditioned media were removed and concentrated using Amicon centrifugation. Proteins in the media were analyzed by Western blot for collagen I, III, and fibronectin using the specific antibodies indicated; (C) Proteins in the media was analyzed by Western blot for MMP1 using the specific antibodies indicated; (D) mRNA expression was examined in HaCaT cells treated with 2 μM DK223 for 24 h using RT-PCR with the specific primers indicated; (E,F) Zymograph assay of the conditioned media from HaCaT cells treated with 2 μM DK223 for 24 h. Conditioned media were removed and concentrated using Amicon centrifugation. Proteins in the media were analyzed by a gelatin zymograph, as described previously. A histogram shows the results of ImageJ data analysis. Data are represented as the mean percentage of distance ± SD fromat least three replicates, * p < 0.05 in all under experiments.