Figure 2
Defining the CER and PER of ZIKV- and mock-infected cells. (A) Mock-infected ERmoxGFP and Sec61β-GFP stably transfected U87 cells were imaged by 3D STED microscopy. Magnified ROIs (yellow ROIs identified by red Roman numerals) show that the PER extends over 3–5 sections (210 nm step size) and CER > 10 sections. Graph shows average PER and CER Z-height for each ERmoxGFP or Sec61β-GFP labeled cell. A Z-height cutoff of 1.26 microns (red line) was used to identify PER and CER objects. (B) Segmented ER labeling from 48 h ZIKV- or mock-infected ERmoxGFP or Sec61β-GFP stably transfected U87 cells (MOI = 1) was visualized using a Z-height spectrum heat map and CER (green; > 1.26 μm) and PER (red; < 1.26 μm) masks are shown. (C) Volume percentage (left) and mean normalized density (right) of CER and PER masks between mock- and ZIKV-infected cells. 5 cells per biological replicate were analyzed for a total of N = 3. ANOVA with post-hoc Tukey HSD: * = P < 0.05, ** = P < 0.01, and *** = P < 0.001. Error bars represent SEM. Scale bar: 10 microns.
