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Figure 3


Fig. 3. siRNA blockade of ß5-subunit expression blocks syndecan-induced cell spreading. (A) Suspended cells are analyzed by flow-cytometry with antibodies capable of recognizing mouse ß1 (HMß1-1), ß3 (2C9.G2) or {alpha}v (H9.2B8) integrin subunits, mAb 281.2 specific for mouse Sdc1, or nonspecific IgG control (gray fill). Cells treated with ß5-integrin-specific or control siRNA are compared. (B) Representative western blot of lysates of cells treated with 0, 200, 400, 600 or 800 nM ß5-specific siRNA and probed for expression of ß5-integrin subunit. FAK expression levels are shown as a loading control. (C) Quantification (± s.e.) of relative ß5 integrin subunit expression from duplicate blots as described in (B). (D) B82L cells were plated on wells coated with 60 µg/ml mAb 281.2 and increasing amounts of VN after treatment with ß5-integrin-specific or control siRNA. Cells were allowed to spread 2 hours before fixation and labeling with Rhodamine-phalloidin. Bar, 50 µm.





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