Fig. 2
PBA treatment resulted in relocalization of PCA267T from the ER and Golgi to GRASP55-positive vesicles. Confocal images from PC (green) and PDI (red)-stained (A), PC (green) and GM130 (red)-stained (B), or PC (green) and GRASP55 (red)-stained (D) CHO-K1 cells stably transfected with PC-A267T untreated (A, B, a–c) or treated with 5 mM PBA for 48 h (A, B, d–f, D, a–c). A Cells were stained with rabbit polyclonal anti-PC (a, d), and mouse monoclonal anti-PDI (b, e), and merged images of green and red are shown in c, f. Co-localized green and red pixels are shown in yellow color. B Cells were stained with rabbit polyclonal anti-PC (a, d) and mouse monoclonal anti-GM130 (b, e), and merged images are shown in c, f. Three independent experiments were performed. Bar 20 µm. C The co-localization of PC with PDI or GM130 was calculated based on the merged (c, f) images in A and B, by the Manders’ co-localization coefficient. Results are presented statistically as the mean ± SEM of at least three independent experiments. *p < 0.05 Students’ t test comparing PBA-treated cells relative to non-treated (ctrl) cells in the PC/PDI-stained samples. **p < 0.05 Students’ t test comparing PBA-treated cells relative to non-treated (ctrl) cells in PC/GM130-stained samples. D Cells were stained with rabbit polyclonal anti-PC (a), and mouse monoclonal anti-GRASP55 (b). Merged images of a and b are shown in c, with co-localized green and red pixels shown in yellow color. Zoomed images of the vesicles are shown in a′–c′. Three independent experiments were performed. Bar in a–c 5 µm. Bar in a′–c′ 2 µm