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. 2015 Feb 5;96(2):266–274. doi: 10.1016/j.ajhg.2014.11.019

Figure 4.

Figure 4

Potential Cytotoxicity of Altered DDX58 in HTM cells

(A) Phase-contrast photographs of HTM cells transduced with empty vector (null) or expression vectors for the WT, two DDX58 alterations (p.Cys268Phe and p.Glu373Ala), and two other known SNPs (rs11795404 [c.548G>T (p.Ser183Ile)] and rs951618 [c.1217T>C (p.Ile406Thr)]) are shown. The scale bar represents 100 μm.

(B) HTM cells were incubated with anti-FLAG antibody diluted 200× or Cy3-conjugated anti-vimentin antibody diluted 500×. The cells were washed and reacted for 2 hr with FITC-conjugated secondary antibody. After the nuclei were counterstained for 20 min with DAPI, the cells were viewed under a fluorescence microscope with the appropriate filter sets and a 200× objective. FITC fluorescence (green) of DDX58 staining merged with DAPI (blue) is shown in the left column, and Cy3 fluorescence (red) of vimentin staining merged with DAPI is shown in the right column. The scale bar represents 100 μm.

(C) Cell viability (%) was calculated as the percentage of cells surviving in comparison to the null. The data were statistically analyzed by one-way ANOVA followed by the Newman-Keuls multiple-comparison test, and p < 0.05 was considered significant. The data are presented as the mean percentage ± the SEM (∗p < 0.01 versus WT).