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. Author manuscript; available in PMC: 2018 Aug 1.
Published in final edited form as: Exp Eye Res. 2017 May 30;161:17–29. doi: 10.1016/j.exer.2017.05.011

Figure 2. Correlation of quantitative assay results with morphology of 661W cells treated with Stsp.

Figure 2

Selected phase-contrast images of cells following the assays described in Fig. 1, treated as follows: A,C,E,G,I, and K: CaAM assay included probenecid; B,D,F,H,J, and L: CaAM assay without probenecid. A,B: VC; C,D: Stsp at 5 nM; E,F: Stsp at 25 nM; G,H: Stsp at 125 nM; I,J: Stsp at 500 nM; K,L: Stsp at 1000 nM. As the Stsp dose increased, the following changes were noted in the appearance of the cultured cells, and with good correlation with the results of the viability assays depicted in Fig. 1: Rounding up of cells (arrows in E,F); Membrane blebbing (arrows in G,H); Contracted aggregates of cells (arrows in I,J); detachment and loss of cells from the substrate (arrows in K,L); Cell shrinkage and collapse of contents, resulting in small phase-bright spheres (arrowheads in I,J,K,L). Cells that underwent either VC or 5 nM Stsp treatment did not exhibit changes indicative of loss of viability, and did show areas of cell overgrowth, suggesting continued proliferation during the course of the experiment (short arrows in A,B,C,D). Magnification bar = 100 μm; insets: 2× magnification of main image; all images for each treatment were captured with a 10× objective lens with phase-contrast optics.