Figure S1

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Figure S1. Effect of physical confinement on α4Ser988 phosphorylation. (A) Schematic diagram of the microchannel device attached to a glass slide. A close-up ...
Cell Reports, Volume 15

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Confinement Sensing and Signal Optimization via Piezo1/PKA and Myosin II Pathways Wei-Chien Hung, Jessica R. Yang, Christopher L. Yankaskas, Bin Sheng Wong, Pei-Hsun Wu, Carlos Pardo-Pastor, Selma A. Serra, Meng-Jung Chiang, Zhizhan Gu, Denis Wirtz, Miguel A. Valverde, Joy T. Yang, Jin Zhang, and Konstantinos Konstantopoulos



Figure S1. Effect of physical confinement on α4Ser988 phosphorylation. (A) Schematic diagram of the microchannel device attached to a glass slide. A close-up detail of channel array is also shown. The red arrow (left to the close-up channel array) indicates the direction of cell migration. (B) CHO-α4WT and CHO-α4S988A cells were seeded in the cell seeding area (unconfined) and then induced to migrate through 20-µm, 10-µm, 6-µm, or 3-µm channels. Cells were imaged by dual-color confocal microcopy for anti-phospho-α4Ser988 (red), GFP (green) or both (merge). (D) Schematic diagram of microcontact printing procedure. (a and b) PDMS stamp (blue) was coated with fibronectin (orange) to be printed. (c) Fibronectin was transferred by printing onto a cover slide (gray) and then examined by fluorescence staining (d). (E) CHO-α4WT and CHO-α4S988A cells placed on either 2D or 1D fibronectin-printed cover slide were imaged by dual-color confocal microcopy for anti-phospho-α4Ser988 (red), GFP (green) or both (merge). (C and F) For each condition, the relative level of α4Ser988 phosphorylation was calculated from the intensity of phospho-α4Ser988 (red fluorescence) normalized to the intensity of α4-GFP (total α4 integrin, green fluorescence). Data represent the mean±SEM. *, p