Fig. 4

Immunofluorescence staining of cells cultured on various nanoporous stainless steel artificial microenvironments. Cells were seeded on different artificial microenvironments and harvested after days 1 or 3. Cytoskeletal arrangement was examined using phalloidin, and vinculin localization was examined using anti-vinculin antibodies (green dots or green-stained area). After day 1, cells displayed an extended morphology on 100-nm nanoporous surfaces, which transitioned to an abnormal elongated or shrunken morphology on 220-nm nanoporous surfaces. A plethora of focal adhesions were seen in cells on flat, 100-nm nanoporous surfaces as compared to 200- and 220-nm nanoporous surfaces. Arrows represent the green spots, i.e., focal adhesions. A similar trend in cell morphology and the number of focal adhesions was seen after day 3. All images were taken at 400×. Scale bars = 50 μm