Fig. 4

Effect of Aqp1 expression on specific cell functions. a, Representative images showing the engulfment of silica beads coated with a supported lipid bilayer (atto390, magenta) by Aqp1-IRES-GFP and GFP-expressing J774A.1 cells (GFP, green). Images were collected using a spinning disk confocal microscope with a 40 × 0.95 NA Plan Apo air objective. Yellow arrows denote beads engulfed by the cells. Scale bar is 5 μm. b, Phagocytic activity of Aqp1- and GFP-cells following exposure to lipid-coated silica beads with or without IgG1κ incorporated in the supported lipid bilayer, for 45 min. Phagocytic index was determined by measuring the average att390 fluorescence per macrophage.c, Glucose-stimulated insulin secretion in Aqp1- and GFP-expressing MIN6 cells induced by exposing cells to 20 mM external glucose for 1 h. d, Flow cytometry analysis of CD25 and CD3 surface expression in Aqp1- and e, GFP-expressing Jurkat cells, either untreated or incubated with anti-human CD3/CD28 beads at a 4:1 bead-to-cell ratio for 24 h. The numbers in each quadrant denote the fraction of cells showing surface expression of CD3 (pan T-cell marker), CD25 (activation marker), both CD3 and CD25, or neither marker. f, Percentage of CD25⁺ cells relative to the total CD3⁺ population. g, Representative images showing the migration of Aqp1- and GFP-expressing MDA-MB-231 cells beyond the Matrigel perimeter on day 6. Wide-field images covering both the Matrigel drop and the surrounding media were acquired using a scanning confocal microscope with a 10 × 10 grid of 1024 × 1024 pixel tiles at 10X magnification. h, Increase in the total area occupied by invading MDA-MB-231 cells that migrate out of the Matrigel. Error bars represent the s.e.m. from n ≥ 3 biological replicates. * P-value < 0.05, ** P-value < 0.01, *** P-value < 0.001 (2-sided, t-test)