Figure 5.

The 3’UTR of GSN mRNA is sufficient for the mRNA localization in the proximity of the plasma membrane. (A-D) Immunofluorescence confocal microscopy and corresponding quantification analyses of EGFP subcellular localization in H1299 (A, B) and A549 (C, D) cells transfected with EGFP-vector and EGFP-3’UTR plasmids. (E-H) Immunofluorescence confocal microscopy and corresponding quantification analyses of endogenous GSN subcellular localization in H1299 (E, F) and A549 (G, H) cells at various confluences. Cells were seeded at 1 × 10⁴ cells/cm² and cultured for a total of 24, 48 or 72 h in standard culture medium. At the indicated confluence, cells were fixed and immunolabelled with antibodies for detection of GSN. DAPI was used for nuclear staining. F-actin was stained using rhodamine-conjugated phalloidin. Arrows: endogenous GSN proteins at the proximity of the plasma membrane. (I, J) Immunofluorescence analyses of the localization of transfected EGFP and endogenous GSN proteins in H1299 cells transfected with the indicated plasmids. Quantification analysis was performed based on counting of cells with diverse protein distribution patterns from three independent experiments (> 15 cells each, n=3). Scale bars: 10 μm.