Fig. 4

HDACi upregulate heparanase mRNA and protein. a Analysis of HPSE expression by qRT-PCR in SS cells treated with SAHA or FK228 for the indicated times. Data are reported as relative quantification with respect to untreated cells as calibration sample. Relative mRNA values are the mean ± SE from three independent experiments. b Western blot detection of heparanase polypeptides in CME-1 cells exposed to SAHA or FK228 for the indicated times. Image on the right is from a cropped blot (dashed line) from which lanes not of interest have been removed. c N-glycosylation inhibition does not modify electrophoretic mobility of heparanase polypeptides. CME-1 cells were treated with tunicamycin (2 μg/ml for 24 h) and processed for Western blotting. As controls, the shift of PDGFRα bands shows the glycosylation inhibition and BIP upregulation indicates endoplasmic reticulum stress. In (b) and (c) anti-actin, −vinculin and -GAPDH blots are shown as loading control