Fig 1.
Workflow and confocal microscopy images comparing WGA membrane staining before vs. after fixation.
Each microscopy image is paired with a schematic illustrating the WGA membrane staining approach. Nuclei were counterstained with DAPI (blue), and cell membranes and organelles were labeled with WGA conjugated to Alexa Fluor 647 (purple). Scale bar: 10 µm. A) WGA staining after fixation showed increased cytoplasmic staining. B) WGA staining before fixation showed clear definition of the plasma membrane, with minimal cytoplasmic staining.
Fig 2.
Confocal microscopy images comparing Gag and Nef (clones EH1 and 3D12) expression in control samples.
A & B) For the confocal images, the stained nuclei (blue) and the plasma membrane (purple) were used to identify the cytoplasmic and membrane region, thus enabling the localization of Gag (green) and Nef (red) protein expression within the cells. Scale bar: 5 µm. A) Dual screening with Nef (EH1) and Gag antibodies. B) Dual screening with Nef (3D12) and Gag antibodies. C) Nef (EH1) and Gag antibodies showed a less correlation coefficient, while D) Nef (3D12) and Gag antibodies showed a high correlation coefficient. Colocalization analyses were performed across the entire regions shown in the images, using three independent confocal images. The colocalization coefficients (Rcoloc) are presented as mean ± SD (n = 3). Statistical significance was assessed using a student’s t-test (p = 0.0151).
Fig 3.
Confocal microscopy images comparing Gag and Nef (clones EH1 and 3D12) expression in H9MN and H9MN-FD cells.
For the confocal images, the following cellular components were stained: the nuclei (blue), Gag protein (green), Nef protein (red) and the plasma membrane (purple). Scale bar: 5 µm. A) Dual screening with Nef (EH1) and Gag antibodies. B) Dual screening with Nef (3D12) and Gag antibodies.