Fig. 4

PPP2R5D and NS5B showed colocalization in the ER of Huh7.5 cells. Localization of PPP2R5D, NS5B, and ER were analyzed by confocal microscopy. A Huh7.5 cells were transfected with plasmids expressing HA-PPP2R5D and Flag-NS5B for 36 h, and then the cells were fixed and immunostained with anti-HA and anti-Flag primary antibodies plus corresponding secondary antibodies using channels 568 nm (indicated by red color) and 488 nm (green), respectively. The nucleus was stained by DAPI. Scale bar, 10 µm. B The colocalization of Flag-PPP2R5D and HA-NS5B was measured by fluorescence intensity line measurement (indicated by an arrow in the merged image in panel A). C Huh7.5 cells were transfected with plasmids expressing HA-PPP2R5D, Flag-NS5B, and DsRed2-ER for 36 h, and then the cells were immunostained and visualized using channels 647 nm (HA-PPP2R5D, red), 488 nm (Flag-NS5B, green), and 568 nm (DsRed2-ER, purple). Scale bar, 5 µm. D The colocalization of Flag-PPP2R5D, HA-NS5B, and ER was measured by fluorescence intensity line measurement (the arrow in the merged image in panel C)