File S1 - Human Equilibrative Nucleoside Transporter-1 Knockdown Tunes Cellular Mechanics through Epithelial-Mesenchymal Transition in Pancreatic Cancer Cells

Figure S1, Western blots of hENT1 (55 kDa) and GAPDH (37 kDa) in pancreatic cancer cells. Figure S2, Young's modulus of Panc 03.27 cells at different indentation force from 10 pN to 10000 pN (inset: Bright-field image showing AFM tip approaching cells and schematic diagram of indentation of microparticle-modified cantilever and cell) (A), Calculated average indentation depth (nm) corresponding to Figure S2A using Eq. (1), and representative force-displacement (f-d) curve from a Panc 03.27 control cell when the indentation force is (C) 100 pN and (D) 1 nN (red: approach curve, black: retract curve) (B). The calculated Young's moduli from (C) and (D) are 1.88 and 5.21 kPa, respectively. Figure S3, Stiffness distribution of (A) Capan-1 and (B) Panc 03.27 cells corresponding to bar histograms shown in Figure 1. The solid line shows Lorentizan distribution. Figure S4, Cellular stiffness of Panc 03.27 cells: Ctrl (without treatment); WGA treated (cell membrane is stained by Alexa Fluor 488 Conjugated wheat germ agglutinin). Young's modulus of cells measured by AFM under same indentation force at 100 pN. Figure S5, Representative confocal micrographs of pancreatic cancer Capan-1 and Panc 03.27 cells showing cytokeratin 18 (green, top panel), Lamin A/C (green, middle and bottom panels), and nuclei (blue) (A). (B) Western blots of Lamin A/C (74, 63 kDa), cytoketarin 18 (46 kDa) and GAPDH (37 kDa) in control, scramble siRNA transfected, and hENT1 knockdown Capan-1 and Panc 03.27 cells. Figure S6, Western blots of E-cadherin (110 kDa), N-cadherin (140 kDa), vimentin (57 kDa), and GAPDH (37 kDa) in untreated and TGF-β treated Panc 03.27 cells (A), (B) Young's modulus of untreated and TGF-β treated Panc 03. 27 cells (concentration of TGF-β: 10 ng/ml, exposure for 2 days). Figure S7, Representative AFM topographic image, deflection image, and corresponding stiffness map of Panc 03.27 cells obtained by using sharp MSCT-C (A, B) and microparticle modified cantilever (D, E). Histograms show (C, F) corresponding stiffness from force volume map (B, E). Figure S8, Young's modulus of cells measured by AFM under same indentation force at 100 pN (A), (B) Western blots of E-cadherin (110 kDa), vimentin (57 kDa), and GAPDH (37 kDa) expressed in six different pancreatic cancer cells. (DOCX)