LARP1 relocalizes to the ER upon HCV infection.

Cells were infected by HCV for 4 d prior to fixation, LARP1, NS5A, and calnexin staining, and confocal microscopy. A. Li diagrams and Li coefficient (B) calculations for LARP1 and Calnexin expression. Pixels present on the left and right sides of the y-axis, i.e., associated with negative and positive staining amplitude values, indicate exclusion and colocalization, respectively. Li coefficients were calculated using the JACop plugin from the ImageJ software (http://rsb.info.nih.gov/ij/plugins/track/jacop.html). Twelve to 15 random fields were acquired per biological sample, totaling 250–300 cells analyzed for each biological sample in a given single experiment (n = 3). C. Representative immunofluorescence-based localization of LARP1, HCV core, and calnexin. Nuclei were counterstained with Hoechst 33342. LARP1 (red), core (magenta), and calnexin (green) were detected using Alexa-617, Alexa-594, and Alexa-488, respectively. Overlays were generated by the ImageJ software. Open and solid arrows show no or total colocalization, respectively. Bar = 5 μm. D. Statistical assessment of the colocalization of LARP1 and Calnexin. Red (LARP1) and green (Calnexin) fluorescence intensities were measured for each pixel along a 5-μm horizontal line centered around the arrow tips in (C) using the Plot Profile function of the ImageJ software. Spearman correlation coefficients for each couple of intensity values are shown. The underlying data for panels in this figure can be found in S1 Data.