Cells were fixed with 4% paraformaldehyde for 5 min, rinsed in
1X phosphate buffered saline, incubated within 0.1% Triton X-100 solution
for 5 min, rinsed, and blocked by a 1% bovine serum albumin solution
(Millipore Sigma) for 30 min. Focal adhesions were visualized using
antibodies specific for vinculin (MAB3574; Millipore Sigma) and integrin
αvβ3 (MAB1976Z; Millpore Sigma). Fibrillar adhesions were
identified using specific antibodies to tensin-1 (NBP1–84129;
Novus biologicals, Toronto, ON), integrin β1 (MAB17781; R&D
Systems, Minneapolis, MN), and integrin α5 (ab150361; Abcam,
Waltham, MA). Extracellular matrix deposition was identified using
antibodies to fibronectin (ab1954; Abcam, Waltham, MA). F-actin was
labeled with rhodamine conjugated phalloidin (R415; Life Technologies,
Grand Island, New York), and nuclei were counterstained with Hoechst
(H3560; Invitrogen, Burlington, ON). Images were taken on a Carl Zeiss
Axio Imager M2m microscope (Zeiss Microscopes, North York, ON) under
reverse osmosis water immersion and processed using Zen Pro software.
1X phosphate buffered saline, incubated within 0.1% Triton X-100 solution
for 5 min, rinsed, and blocked by a 1% bovine serum albumin solution
(Millipore Sigma) for 30 min. Focal adhesions were visualized using
antibodies specific for vinculin (MAB3574; Millipore Sigma) and integrin
αvβ3 (MAB1976Z; Millpore Sigma). Fibrillar adhesions were
identified using specific antibodies to tensin-1 (NBP1–84129;
Novus biologicals, Toronto, ON), integrin β1 (MAB17781; R&D
Systems, Minneapolis, MN), and integrin α5 (ab150361; Abcam,
Waltham, MA). Extracellular matrix deposition was identified using
antibodies to fibronectin (ab1954; Abcam, Waltham, MA). F-actin was
labeled with rhodamine conjugated phalloidin (R415; Life Technologies,
Grand Island, New York), and nuclei were counterstained with Hoechst
(H3560; Invitrogen, Burlington, ON). Images were taken on a Carl Zeiss
Axio Imager M2m microscope (Zeiss Microscopes, North York, ON) under
reverse osmosis water immersion and processed using Zen Pro software.