Fig 4. Co-culture of hMSCs transduced with ADAMTS5-targeting shRNA effectively reduces ADAMTS5 expression in flow cytometry sorted RFP-labeled SW982.

(A) Following co-culture of non-targeting control shRNA or ADAMTS5-targeting shRNA transduced hMSCs with RFP labeled SW982 cells, the cells were sorted by flow cytometry. The gating threshold (RFP+ sorted) was set based on the analysis of co-cultured hMSCs with unlabeled SW982 cells (no SW982-RFP cells). RFP-positive SW982 cells that were co-cultured with hMSCs previously transduced with non-targeting control-shRNA or ADAMTS5-targeting shRNA were sorted directly into TriPure reagent for RNA isolation. (B) The histogram shows the relative gene expression for ADAMTS5 in the RFP-positive cell population isolated after co-culture with hMSC-shRNA-control or hMSC-shRNA-ADAMTS5 cells as determined by quantitative real time RT-PCR. The result is from a representative experiment.