Fig. S1. β3 integrin, but not β5 integrin, mediates IF-FA association. (A) Surface expression of αvβ3, αv, and β5 integrins was evaluated by FACs using untransfected TrHBMECs or TrHBMECs transfected with β3 integrin siRNA (2 ab=non-transfected TrHBMECs labeled with secondary antibody only). (B) TrHBMECs transfected with β3 integrin siRNA were stained for paxillin (green) and actin (red) or paxillin (green) and tubulin (red) and overlays of the green and red channels are presented. (C) TrHBMECs transfected with control siRNA or β5 integrin siRNA were stained for β5 integrin (green) and vimentin (red) as indicated. From left to right, the first panel shows an overlay of the green and red channels. The second panel shows the lack of β5 integrin at FA-like structures. However, the vimentin remains well distributed throughout the cell as can be seen in the third panel and the phase image. Scale bars: 10 m.

Fig. S2. The actin and microtubule networks are normal in TrHBMECs spread on PLL. TrHBMECs were plated in serum-free medium on PLL-coated coverslips overnight. Cells were processed for immunofluorescence microscopy using a combination of antibodies against actin (red) and paxillin (green) or tubulin (red) and paxillin (green) and overlays of the green and red channels are presented. The boxed areas are shown at higher magnification in the insets. Scale bars: 10 m.

Fig. S3. The microtubule network in TrHBMECs remains intact after treatment with dynein or kinesin shRNAs. TrHBMECs electroporated with heavy chain dynein or heavy chain kinesin shRNA were plated in serum-free medium on PLL-coated coverslips overnight. Untreated cells (left panels) or cells incubated for 30 minutes in medium supplemented with 0.5 mM MnCl₂ (right panels) were processed for immunofluorescence microscopy using a combination of antibodies against paxillin (green) and tubulin (red). Overlays of the green and red channels are presented. Scale bars: 10 m.

Fig. S4. Plectin siRNA does not disrupt the actin or microtubule networks in TrHBMECs. Mock transfected (control) TrHBMECs or TrHBMECs transfected with plectin siRNA were stained for paxillin (green) and actin (red) or paxillin (green) and tubulin (red) and overlays of the green and red channels are presented. Scale bars: 10 m.