B. MAb 9E1 reduces MMP-9 activity in MDA-MB-231 cells. A

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B. MAb 9E1 reduces MMP-9 activity in MDA-MB-231 cells. A Figure 1: Characterisation of MAb 9E1. A. MAb 9E1 reacts with a 65 kDa reactive band A 65Kda reactive band is identified across breast cancer cell lines. BT-20 does not express the MAb target antigen and reduced expression is observed in MDA- MB-468 (absent in membrane enriched fractions). Reactivity is observed in both total cell lysates and membrane enriched fractions. Duplicate biological samples were probed with MAb 9E1, a representative immunoblot is shown. B. MAb 9E1 reduces MMP-9 activity in MDA-MB-231 cells. (i) Gelatin zymography of conditioned medium from MDA-MB-231 cells with intense clear zone showing active MMP-9 (92 kDa). A very weak signal indicative of pro MMP-9 can be observed directly underneath the active form. (ii) Representative gelatin zymogram showing analysis of conditioned medium (n=2). following treatment of cells with 50ug/ml of MAb 9E1. A reduction in MMP-9 activity is observed in antibody treated conditioned medium compared relative to control hybridoma medium only (no antibody). Densitometry analysis of reduced MMP-9 activity (fold change) is also shown. . A Whole cell lysates Membrane enriched fractions Alpha tubulin B i ii 110 kDa 80 kDa active MMP-9 110 KDa active MMP-9 Fold change Hybridoma medium control MAb 9E1 (no antibody)