Figure 8

Zymography analysis for active MMP2 was performed with 4 μg of total protein of the conditioned medium of U87 (a) and primary IC cells (b) after 48 h exposure to JS-K (1–3 μM) on gelatin-containing PAGE. Western blot analysis for MMP2, MMP7, MMP9 and TIMP3 was performed for conditioned medium of U87 (a) and IC (b) cells with 4 μg of total protein as well as of 20 μg of total cell lysate of U87 (c) and IC cells (d) on SDS-PAGE. Conditioned media of knockdown and overexpression of ATF3 were examined for active MMP2 as well as the plasmid controls and uninfected U87 (e). 4 μg of protein of the supernatant (e) and 20 μg of cell lysates (f) were screened for MMP2, MMP7, MMP9 and TIMP3 by SDS-PAGE. Coomassie staining of the gels was used as a loading control for zymogram and for western blot of the supernatant, GAPDH was used as a loading control for cell lysates. Figures are representative for three independent experiments.