Figure 2
Differential expression of ATX in human liver cancer cell lines (Huh7, Hep3B and HepG2), normal embryonic liver cell line CL-48, and normal primary hepatocytes. A. Comparison of ATX mRNA levels by qRT-PCR analysis. Columns are average ± SD from three independent experiments. B. Equal number of cells (4 × 105) were plated into 100 mm dishes and incubated in serum free EMEM containing 0.1% BSA for 24 hours. Cells were lysed with RIPA buffer and 15 μg of lysates was used for SDS-PAGE and probed for ATX antibody, β-actin level was used as a loading control (top two pannels). 1/10 volume of concentrated medium was used for immunoblot (bottom panel). An intense band of 110 kDa was detected in the cell lysate from Hep3B and Huh7 cells, which is corresponding to the positive ATX control. The sizes of the MW markers are shown on the right. rATX indicates recombinant ATX protein which is a positive control. IB indicates immunoblot.