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. 2021 Dec 5;13(23):6124. doi: 10.3390/cancers13236124

Figure 5.

Figure 5

Subcellular localization of edelfosine in the ER of PANC-1 cells and PANC-1 CSC pancreatic cancer spheroids, and induction of an ER stress response. (a) Live PANC-1 cells and PANC-1 CSC pancreatic cancer spheroids were labeled overnight at 37 °C for the ER in red using the CellLight ER-RFP BacMam 2.0 reagent, and then the samples were incubated with 20 μM Et-BDP-ET (green fluorescence) for 3 h at 37 °C. Areas of colocalization between the ER and Et-BDP-ET in the merge panels are yellow. The cells were also stained for nuclei with DAPI (blue fluorescence). The corresponding differential interference contrast (DIC) microscopy images are also shown. Bar, 25 μm. The data are representative of three independent experiments. (b) PANC-1 cells and PANC-1 CSCs were untreated (C) or treated with 20 μM edelfosine for the indicated times and analyzed by means of Western blotting using specific antibodies for the denoted proteins; β-actin was used as the loading control. Molecular weights (in kilodaltons) of every protein are indicated at the right side of each panel. The gels were cropped to show the relevant sections. Relative protein level quantification after normalization to the internal control β-actin is included below each Western blot panel. The Western blot images are representative of three independent experiments.